KR20040039422A - Particulate hydrophobic polymer, production process therefor and column for reversed-phase high-performance liquid chromatography - Google Patents

Abstract

After reacting the granular crosslinked polymer (A) having a hydroxyl group with the crosslinkable epoxy compound (B) and hydrolyzing the epoxy bond, the compound having the obtained hydroxyl group and the epoxy compound (C) having 6 to 40 carbon atoms in total are reacted. A granular hydrophobic polymer produced by the method, a production method thereof, a column for reverse phase liquid chromatography, and an analysis method using the column. By using the granular hydrophobic polymer of the present invention with remarkably high acid / alkali resistance, the swelling and shrinkage of the granular polymer itself is reduced, so that even if the eluents in the column are variously exchanged, the column efficiency in various solvents is not lowered and the polycyclic aromatic Sharp chromatograms of compounds can be obtained.

Description

Granular hydrophobic polymer, preparation method thereof, and column for reverse phase high performance liquid chromatography {PARTICULATE HYDROPHOBIC POLYMER, PRODUCTION PROCESS THEREFOR AND COLUMN FOR REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY}

As one separation mode of liquid chromatography, reverse phase liquid chromatography is widely known in which separation is performed based on different retention strengths due to the difference in hydrophobicity between the filler and the material to be separated.

Conventionally, as a filler for reverse phase liquid chromatography, a filler obtained by introducing an octadecyl group into a silanol group of silica gel using a silane coupling agent having an octadecyl group (hereinafter referred to as "ODS") is widely used. . This is because fine particles having high mechanical strength can be produced relatively easily.

However, since silica gel has low alkali resistance, usable eluents are limited to eluents having a pH of 2-8. In addition, since it is virtually impossible to chemically bond the coupling agent to all silanol groups, it causes a serious problem that basic compounds such as amines are easily adsorbed to unreacted silanol groups.

In recent years, various polymer fillers for reverse phase liquid chromatography have been developed to improve handleability under alkaline conditions, and some of them are commercially available as columns for reverse phase liquid chromatography. Here is an example:

(1) Styrene-Divinylbenzene Copolymer Particles (Analytical Chemistry, Vol. 45, page 1383 (1973), commercially available: Shodex (registered trademark of Showa Denko Co., Ltd.) RSpak RP18- manufactured by Showa Denko Co., Ltd.) 413);

(2) methacrylate-based crosslinked polymer particles (commercially available product: Shodex (registered trademark of Showa Denko Co., Ltd.) RSpak DE-413, available from Showa Denko KK);

(3) Polyvinyl alcohol-based crosslinked polymer particles having a chemically bonded long-chain acyl group (commercially available product: Shodex (registered trademark of Showa Denko Co., Ltd.)) Asahipak (registered trademark of Showa Denko Co., Ltd.) Brand) ODP-40 4D);

(4) methacrylic acid ester copolymer particles having a long chain alkyl group (see JP-A-2000-9707 ("JP-A" as used herein means "Japanese Patent Publication");

(5) copolymer particles of glycidyl methacrylate and a (meth) acrylic acid ester of a polyhydric alcohol having a long chain alkyl group chemically bonded thereto (see JP-A-61-272654); And

(6) Hydroxyl group-containing methacrylate crosslinked polymer particles having chemically bonded long chain acyl groups (see JP-A-4-58154).

The column for reverse phase liquid chromatography packed with these polymer particles is advantageous in that the pH range which is usable is wider than that of the ODS column, but there are also few problems.

However, since the chromatogram of a polycyclic aromatic compound becomes wide, the column filled with the granular polymer of (1)-(5) is not suitable for the separation and analysis of the natural product or medicine which has a polycyclic aromatic site | part.

In the column filled with the granular polymer of (1), it is difficult for the granular polymer itself to swell or shrink seriously depending on the solvent, and to perform the separation and analysis satisfactorily by changing the eluent in the column.

The column filled with the granular polymer of (2) to (6) has a low resistance to acid / alkali because the ester bonds contained in the structure have a low resistance to acid / alkali, so that when the column is used for a long time at a pH of less than 2 or above pH 11, The column efficiency is markedly lowered, the peak shape of basic substances such as amines is worsened, and there is a problem that the measurement cannot be continuously performed.

As a method for solving this problem, it is effective to produce an ether bond having high resistance to acid / alkali by reacting an epoxy compound with a granular polymer having a hydroxyl group upon introduction of a functional group. Examples of the method include (A) a method of carrying out the reaction in water containing a base such as sodium hydroxide or a polar organic solvent such as dimethylformamide, and (B) Lewis such as a boron trifluoride diethyl ether complex. A method of carrying out the reaction in an ether solvent such as dioxane in the presence of an acid (JP-A-61-272654 mentioned in (5) above) is known. However, these methods have a problem that the reaction is carried out for a long time and the post-treatment such as filtration becomes very cumbersome because the epoxy compound is used in an excessive amount. In addition, if the base gel is a polyester-based gel as in the case where the material (5) is used, since the base gel is exposed to an acid or an alkali for a long time, when the reaction of (A) or (B) is carried out, the granular polymer The ester bond of itself is cleaved by hydrolysis, and a carboxyl group appears. The granular polymer having a carboxyl group adsorbs basic substances such as amines, and therefore is not suitable as a filler for reverse phase liquid chromatography.

Fillers which can overcome the above-mentioned problems and a method of manufacturing the same are not known, and their establishment is urgently required.

This application, 35 U.S.C. According to Article 119 (e) (1), 35 U.S.C. Claiming the benefit of the filing date of US Provisional Serial No. 60 / 328,794, filed Oct. 15, 2001 under 111 (b), 35 U.S.C. Based on the provisions of Article 111 (a).

The present invention relates to a particulate hydrophobic polymer having excellent acid / alkali resistance as a filler for reverse phase liquid chromatography; Its production method; Columns for reverse phase liquid chromatography filled with particulate polymer; And an analytical method using the column.

An object of the present invention is a novel manufacturing method, which has a remarkably high acid / alkali resistance, reduces the swelling and shrinkage of the granular polymer itself, so that the column efficiency in various solvents can be maintained even if the eluents in the column are exchanged in various ways, It is to provide a filler for reverse phase liquid chromatography that provides a sharp chromatogram for polycyclic aromatic compounds.

As a result of earnestly examining in order to solve the above-mentioned problem, the present inventors (1) reacted the granular crosslinked polymer (A) which has a hydroxyl group, and a crosslinkable epoxy compound (B), and after apply | coating a base material, hydrolyze an oxirane ring Afterwards, when the granular polymer obtained by reacting with an epoxy compound (C) having 6 to 40 carbon atoms in total is charged to a column used for reverse phase liquid chromatography, it has a remarkably high resistance to acids and alkalis and solvents. Very high resistance to exchange can be obtained, sharp chromatograms can be obtained for polycyclic aromatic compounds, and (2) reaction of crosslinkable epoxy compounds (B) with epoxy compounds having 6 to 40 carbon atoms in total. It was newly found that the reaction proceeds very rapidly when carried out in the presence of a Lewis acid catalyst in this low polar solvent. This object can be achieved by this finding.

More specifically, the present invention relates to the following.

(1) After reacting a granular crosslinked polymer (A) having a hydroxyl group with a crosslinkable epoxy compound (B) and hydrolyzing the oxirane ring, the compound having the obtained hydroxyl group and an epoxy compound having 6 to 40 carbon atoms in total. A granular hydrophobic polymer produced by reacting (C).

(2) In the above (1), the granular crosslinked polymer (A) having a hydroxyl group is a poly (meth) acrylate (I), glycidyl (meth) acrylate (II) of a polyhydric alcohol having at least one hydroxyl group in a molecule thereof. ) And two or more copolymers selected from the group consisting of poly (meth) acrylates (III) of polyhydric alcohols having no hydroxyl group in the molecule, or homopolymers of poly (meth) acrylates (I) of polyhydric alcohols. A granular hydrophobic polymer characterized by the above-mentioned.

(3) In (2), poly (meth) acrylate (I) of a polyhydric alcohol having one or more hydroxyl groups in the molecule is glycerol dimethacrylate, and the glycidyl (meth) acrylate (II) is Poly (meth) acrylate (III) of a polyhydric alcohol which is glycidyl methacrylate and does not have a hydroxyl group in the molecule is a granular hydrophobic polymer, characterized in that alkylene glycol dimethacrylate.

(4) The granular hydrophobic polymer according to the above (3), wherein the alkylene glycol dimethacrylate is ethylene glycol dimethacrylate.

(5) The granular hydrophobic polymer according to the above (1), wherein the crosslinkable epoxy compound (B) is a compound selected from epihalohydrin and an epoxy compound containing two or more oxirane rings.

(6) In the above (5), the epoxy compound containing two or more oxirane rings is ethylene glycol diglycidyl ether, propylene glycol diglycidyl ether, butylene glycol diglycidyl ether, trimethylolpropane A granular hydrophobic polymer characterized by diglycidyl ether, trimethylolpropane triglycidyl ether, pentaerythritol triglycidyl ether, or triglycidyl isocyanurate.

(7) In the above (1), the epoxy compound (C) having 6 to 40 carbon atoms in total is a compound selected from the group consisting of compounds represented by the following general formulas (1) to (4) Granular hydrophobic polymer.

(Where n represents an integer of 4 to 38.)

(Where n represents an integer of 3 to 37.)

(Where n represents an integer of 0 to 32)

(Where n represents an integer of 0 to 31)

(8) The granular hydrophobic polymer according to the above (7), wherein the epoxy compound (C) having 6 to 40 carbon atoms is stearyl glycidyl ether.

(9) The granular hydrophobic polymer according to any one of the above (1) to (8), wherein the average particle diameter is 1 to 2,000 µm.

(10) After reacting the granular crosslinked polymer (A) having a hydroxyl group with the crosslinkable epoxy compound (B), hydrolyzing the oxirane ring, and further reacting with the epoxy compound (C) having 6 to 40 carbon atoms in total. Method for producing a granular hydrophobic polymer, characterized in that.

(11) In the above (10), the granular crosslinked polymer (A) having a hydroxyl group is a poly (meth) acrylate (I) or a glycidyl (meth) acrylate (II) of a polyhydric alcohol having at least one hydroxyl group in a molecule thereof. And at least two copolymers selected from poly (meth) acrylate (III) of a polyhydric alcohol having no hydroxyl group in the molecule, or a homopolymer of poly (meth) acrylate (I) of a polyhydric alcohol. Process for preparing granular hydrophobic polymer.

(12) The method for producing a granular hydrophobic polymer according to the above (10), wherein the crosslinkable epoxy compound (B) is a compound selected from epihalohydrin and an epoxy compound containing two or more oxirane rings. .

(13) In (11), the poly (meth) acrylate (I) of the polyhydric alcohol having one or more hydroxyl groups in the molecule is glycerol dimethacrylate, and the glycidyl (meth) acrylate (II) is The poly (meth) acrylate (III) of the polyhydric alcohol which is glycidyl methacrylate and does not have a hydroxyl group in the said molecule | numerator is a manufacturing method of the granular hydrophobic polymer characterized by the above-mentioned alkylene glycol dimethacrylate.

(14) The method for producing a granular hydrophobic polymer according to the above (13), wherein the alkylene glycol dimethacrylate is ethylene glycol dimethacrylate.

(15) In (12), the epoxy compound containing two or more oxirane rings is ethylene glycol diglycidyl ether, propylene glycol diglycidyl ether, butylene glycol diglycidyl ether, trimethylolpropane A diglycidyl ether, trimethylolpropane triglycidyl ether, pentaerythritol triglycidyl ether, or triglycidyl isocyanurate.

(16) The granular hydrophobic polymer according to the above (10), wherein the epoxy compound (C) having 6 to 40 carbon atoms is a compound selected from compounds represented by the general formulas (1) to (4). Manufacturing method.

(17) The method for producing a granular hydrophobic polymer according to the above (16), wherein the epoxy compound (C) having 6 to 40 carbon atoms in total is stearyl glycidyl ether.

(18) In the above (10), the granular crosslinked polymer (A) having the hydroxyl group and the reaction of the epoxy compound containing two or more oxirane rings are granulated, characterized in that carried out in the presence of Lewis acid in a low polar solvent. Process for preparing hydrophobic polymer.

(19) The method for producing a granular hydrophobic polymer according to the above (10), wherein the reaction with the epoxy compound (C) having 6 to 40 carbon atoms in total is carried out in the presence of Lewis acid in a low polar solvent.

(20) The method for producing a granular hydrophobic polymer according to (18) or (19), wherein the low polar solvent is a hydrocarbon having 5 to 10 carbon atoms in total.

(21) The method for producing a granular hydrophobic polymer according to (18) or (19), wherein the concentration of the Lewis acid is 1 to 70% by mass relative to the granular crosslinked polymer.

(22) The method for producing a granular hydrophobic polymer according to any one of (10) to (21) above, wherein the granular hydrophobic polymer has an average particle diameter of 1 to 2,000 µm.

(23) A column for reverse phase liquid chromatography, wherein the granular hydrophobic polymer according to any one of (1) to (9) is filled.

(24) A method of analyzing a sample containing a polycyclic aromatic compound,

The analysis method characterized by using the column for reverse phase liquid chromatography as described in said (23).

The present invention relates to a particulate hydrophobic polymer having excellent acid / alkali resistance as a filler for reverse phase liquid chromatography; Its production method; Columns for reverse phase liquid chromatography filled with particulate polymer; And an analytical method using the column.

Examples of poly (meth) acrylates (I) of polyhydric alcohols having at least one hydroxyl group in the molecule used in the present invention include glycerol diacrylate, glycerol dimethacrylate, pentaerythritol triacrylate, pentaerythritol di Acrylate, pentaerythritol trimethacrylate, pentaerythritol dimethacrylate, trimethylolpropane diacrylate and trimethylolpropane dimethacrylate are exemplified. Among these, glycerol diacrylate and glycerol dimethacrylate are preferable from the viewpoint of easy availability and economical efficiency. These compounds may be used independently and may be used in combination of 2 or more type among them.

Glycidyl (meth) acrylate (II) is preferably glycidyl methacrylate in view of the chemical strength of the granular crosslinked polymer produced.

Examples of poly (meth) acrylate (III) of polyhydric alcohols having no hydroxyl group in the molecule include ethylene glycol di (meth) acrylate, diethylene glycol di (meth) acrylate, propylene glycol di (meth) acrylate, polyethylene (Meth) acrylates of polyalkylene glycols such as glycol di (meth) acrylate and polypropylene glycol di (meth) acrylate; Glycerol tri (meth) acrylate; And trimethylolpropane tri (meth) acrylate. Among these, ethylene glycol dimethacrylate is preferable from the viewpoint of easy availability and economical efficiency. These compounds may be used independently and may be used in combination of 2 or more type among them.

In the present invention, a granular homopolymer of poly (meth) acrylate (I) of a polyhydric alcohol having at least one hydroxyl group in a molecule for reaction of the granular crosslinked polymer having a hydroxyl group with the crosslinkable epoxy compound (B), or The granular copolymer of poly (meth) acrylate (I) of the polyhydric alcohol and poly (meth) acrylate (III) of the polyhydric alcohol having no hydroxyl group in the molecule may be reacted with an epoxy compound described later as it is. In addition, poly (meth) acrylate (I), glycidyl (meth) acrylate (II) and glycidyl (meth) acrylate (II) of polyhydric alcohols and poly (poly) alcohols having no hydroxyl group in the molecule ( Meta) acrylate (III) or poly (meth) acrylate (I), glycidyl (meth) acrylate (II) and poly (meth) acrylate (III) of a polyhydric alcohol having no hydroxyl group in the molecule After copolymerization, the oxirane ring in the copolymer may be hydrolyzed, followed by reaction with a crosslinkable epoxy compound described later.

In the preparation of the copolymer of poly (meth) acrylate (I) of a polyhydric alcohol and poly (meth) acrylate (III) of a polyhydric alcohol having no hydroxyl group in the molecule, the amount of (I) is 10 mass% or more. It is preferable, More preferably, it is 20 mass% or more. When the ratio of (I) is less than 10% by mass, the granular polymer obtained after the reaction with the epoxy compound (C) having 6 to 40 carbon atoms in total has poor separation performance as a filler for reverse phase liquid chromatography, and in particular, polycyclic The peak of the aromatic compound is widened.

In the preparation of the copolymer of poly (meth) acrylate (I) of polyhydric alcohol and glycidyl (meth) acrylate (II), the amount of (I) used is preferably 30% by mass or more, more preferably It is 50 mass% or more. If the ratio of (I) is less than 30 mass%, the mechanical strength of the granular polymer obtained will fall.

In the preparation of the copolymer of glycidyl (meth) acrylate (II) and poly (meth) acrylate (III) of a polyhydric alcohol having no hydroxyl group in the molecule, the amount of (III) used is from 30 to 90 mass%. It is preferable, More preferably, it is 50-80 mass%. When the ratio of (III) is less than 30% by mass, the mechanical strength of the granular polymer obtained is lowered. On the other hand, when the ratio of (III) exceeds 90% by mass, the epoxy compound (C) having 6 to 40 carbon atoms and The granular polymer obtained after the reaction has poor separation performance as a filler for reverse phase liquid chromatography, and the peak of the polycyclic aromatic compound becomes wider.

In the preparation of the copolymer of poly (meth) acrylate (I) of polyhydric alcohol, glycidyl (meth) acrylate (II) and poly (meth) acrylate (III) of polyhydric alcohol having no hydroxyl group in the molecule As for the total ratio of (I) and (III) used, 30 mass% or more is preferable, More preferably, it is 50 mass% or more. The total ratio of (I) and (III) is less than 30 mass%, and the mechanical strength of the granular polymer obtained falls. At the same time, 10 mass% or more is preferable, and, as for the total ratio of (I) and (II) used, More preferably, it is 20 mass% or more. When the total ratio of (I) and (II) is less than 10% by mass, the granular polymer obtained after the reaction with the epoxy compound (C) having 6 to 40 carbon atoms in total has poor separation performance as a filler for reverse phase liquid chromatography. And the peak of the polycyclic aromatic compound becomes wider.

In the present invention, poly (meth) acrylate (I) of polyhydric alcohol having at least one hydroxyl group in a molecule, glycidyl (meth) acrylate (II) and poly (meth) of polyhydric alcohol having no hydroxyl group in a molecule The acrylate (III) may be subjected to aqueous suspension polymerization in the presence of an organic solvent which is not compatible with water to form a granular crosslinked (co) polymer (hereinafter, (I), (II) and (III) are respectively). "Monomer").

The organic solvent which is not compatible with water used in the present invention is not particularly limited. However, as the organic solvent used has higher affinity with the monomer, the granular crosslinked (co) polymer has a reduced pore diameter and an increased physical strength. In view of this, an organic solvent containing an alcohol having 5 to 12 carbon atoms as a main component such as isoamyl alcohol, 1-hexanol, 1-octanol, 2-ethylhexanol, 1-decanol, and 1-dodecanol Is preferred. These organic solvents may be used alone, or two or more thereof may be used in combination.

The amount of the organic solvent used may be 10 to 300 mass% with respect to the total amount of monomers (I), (II) and (III), but it is 25 to 100 mass% in view of the specific surface area and physical strength of the granular crosslinked (co) polymer. Is preferred.

Examples of suitable polymerization initiators for use in the present invention include organic peroxides such as benzoyl peroxide, dichlorobenzoyl peroxide, dicumyl and lauroyl peroxide, and 2,2'-azobis (isobutyronitrile) and 2,2 Azo compounds, such as'-azobis (2, 4- dimethylvaleronitrile), are illustrated. These compounds may be used independently and may be used in combination of 2 or more type among them. Among these, an azo polymerization initiator is preferable from a viewpoint of easy handling.

The usage-amount of a polymerization initiator is 0.1-4 mass% with respect to the total amount of a monomer, Preferably it is 1-2 mass%. If the amount of the polymerization initiator is less than 0.1% by mass, a very long time is required to complete the polymerization. On the other hand, if the amount of the polymerization initiator is more than 4% by mass, the polymerization proceeds at a high speed, which is not preferable from the viewpoint of safety.

In the present invention, a dispersant such as poorly soluble phosphate or a water-soluble high molecular compound can be added to the aqueous phase. Examples of poorly soluble phosphates include (third) calcium phosphate and magnesium phosphate. Examples of the water-soluble high molecular compound include polyvinyl alcohol, alkyl cellulose, hydroxyalkyl cellulose and carboxyalkyl cellulose.

The dispersant may be a poorly soluble phosphate or a water-soluble high molecular compound, but a water-soluble high molecular compound is preferable because it can be removed by washing under neutral conditions. As for the usage-amount of a water-soluble high molecular compound, 0.01-3 mass% is preferable with respect to water.

Moreover, in order to reduce the solubility in water of the organic solvent which is incompatible with a monomer or water, you may add a water-soluble inorganic salt to an aqueous phase. Examples of water soluble inorganic salts include sodium chloride, calcium chloride and sodium sulfate. These water-soluble inorganic salts may be used alone, or two or more thereof may be used in combination. Although the density | concentration of the salt used is not specifically limited, For example, it is preferable that sodium chloride is used in 0.5-15 mass% with respect to the water used.

Poly (meth) acrylate (I) of polyhydric alcohol having at least one hydroxyl group in the molecule, glycidyl (meth) acrylate (II), poly (meth) acrylate of polyhydric alcohol having no hydroxyl group in the molecule (III) ) And the polymerization initiator are mixed in advance, and the mixture is added and dispersed in an aqueous solution containing a water-soluble inorganic salt. In this case, a disperser such as a homomixer can be used depending on the target particle size.

Although the use amount of water is 1-50 mass times with respect to the total amount of a monomer and an organic solvent, it is preferable that the use amount of water is 2-10 mass times from a viewpoint of stability of a dispersion solution or the time required for filtration in a post process.

When using monomer (II), the glycidyl group of the obtained granular crosslinked (co) polymer is hydrolyzed by an acid. Examples of acid catalysts are sulfuric acid, perchloric acid, hydrochloric acid, toluenesulfonic acid and benzenesulfonic acid. Among these, hydrochloric acid is preferable from the viewpoint of easy handling.

As for the density | concentration of the acid used, 0.01-5N is preferable, More preferably, it is 0.05-2N. If the concentration is less than 0.01N, the reaction does not proceed rapidly. On the other hand, if the concentration is more than 5N, hydrolysis of the ester bond is likely to occur. At the time of performing this reaction, if a solvent contains at least 10 mass% or more of water, you may use together an organic solvent. The organic solvent used is not particularly limited as long as it is compatible with both acid and water and is inert to acid and glycidyl groups. Examples are acetone, methyl ethyl ketone, 1,4-dioxane and tetrahydrofuran.

The amount of the acid solution to be used may be an amount sufficient to immerse the granular crosslinked polymer in the solvent, and may be, for example, equal to or greater than the gel mass. Although reaction conditions can be arbitrarily determined, it is preferable that reaction is performed at 25-100 degreeC for 3 to 10 hours.

The granular crosslinked (co) polymer thus obtained is surface crosslinked with a crosslinkable epoxy compound (B). This manipulation is necessary to give the particulate polymer sufficiently high mechanical strength and very high acid / alkali resistance.

Examples of the crosslinkable epoxy compound (B) include compounds selected from the group consisting of epihalohydrin and epoxy compounds having two or more oxirane rings. Examples of epihalohydrin are epichlorohydrin, epibromohydrin and epiiodhydrin. Examples of the epoxy compound having two or more oxirane rings include ethylene glycol diglycidyl ether, propylene glycol diglycidyl ether, butylene glycol diglycidyl ether, trimethylolpropane diglycidyl ether, trimethylolpropane Triglycidyl ether, pentaerythritol triglycidyl ether and triglycidyl isocyanurate are exemplified. These compounds may be used independently and may be used in combination of 2 or more type among them. Among these, epichlorohydrin or ethylene glycol diglycidyl ether is preferable in view of the balance between the strength and economical efficiency achieved.

In the case of performing the surface crosslinking reaction using epihalohydrin as the crosslinkable epoxy compound (B), the reaction proceeds while removing or removing the halogen atoms, so that the reaction is carried out in the presence of an aqueous alkali solution. Examples of alkalis commonly used include hydroxides and carbonates of alkali metals, and hydroxides and carbonates of alkaline earth metals. This reaction may be carried out in a mixed solvent with water and a polar organic solvent such as dimethylformamide, N-methylpyrrolidone and dimethyl sulfoxide.

As for the usage-amount of epihalohydrin, 5-200 mass% is preferable with respect to a granular crosslinked polymer, More preferably, it is 50-150 mass%. When the amount of epihalohydrin is less than 5% by mass, the surface crosslinking reaction proceeds at a low speed and takes a very long time. On the other hand, when the amount of epihalohydrin is more than 200% by mass, the post-treatment after the reaction is performed. It becomes very cumbersome.

5-40 mass% is preferable, and, as for the density | concentration of the aqueous alkali solution used, 10-30 mass% is more preferable from a viewpoint of easy manufacture. The amount of the aqueous alkali solution may be any amount sufficient to deposit the particulate polymer in the solvent, for example, two times or more of the gel mass. When the reaction is carried out by adding a polar organic solvent such as dimethylformamide, N-methylpyrrolidone or dimethyl sulfoxide, the polar organic solvent is added to the system in which the aqueous alkali solution is added in an amount of two times or more the gel mass.

Although the addition amount of a polar organic solvent is not specifically limited, You may add in the quantity of 0.1-2 times of aqueous alkali solution. The reaction is preferably carried out at 25 to 100 ° C. for 3 to 16 hours, more preferably at 30 to 60 ° C. for 5 to 12 hours.

When using an epoxy compound having two or more oxirane rings as the crosslinkable epoxy compound (B), the surface crosslinking reaction can be performed under alkaline conditions or acidic conditions. When performing the surface crosslinking reaction under alkaline conditions, the reaction conditions may be exactly the same as when epihalohydrin is used as the crosslinkable epoxy compound (B).

As a method of performing the surface crosslinking reaction under acidic conditions, a method of performing the reaction in the presence of Lewis acid as a catalyst may be used. The Lewis acid is not particularly limited, but examples thereof include ether complexes of boron trifluoride, tin tetrachloride and titanium tetrachloride. These compounds may be used in combination of two or more thereof. Among them, the ether complex of boron trifluoride is most preferable from the viewpoint of easy handling.

Examples of the solvent used in performing the surface crosslinking include benzene, toluene, o-xylene, m-xylene, p-xylene, mesitylene, ethylbenzene, propylbenzene, cumene, butylbenzene, isobutylbenzene, amylbenzene, Isoamylbenzene, pentane, hexane, cyclohexane, cyclopentane, decalin, heptane, octane, isooctane, n-nonane, n-decane, n-undecane, n-dodecane, n-tridecane, n-tetradecane hydrocarbon solvents having 5 to 18 carbon atoms such as n-hexadecane and n-octadecane; Anisole, ethyl isoamyl ether, ethyl-t-butyl ether, diisoamyl ether, diisopropyl ether, diphenyl ether, dibutyl ether, dipropyl ether, dibenzyl ether, tetrahydrofuran, methyl-t- Ether solvents such as butyl ether, 1,4-dioxane, diethylene glycol diethyl ether and triethylene glycol dimethyl; And chloroform, carbon tetrachloride, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, chlorobenzene, o-chlorotoluene, p-chlorotoluene, o-dichlorobenzene, m-dichlorobenzene, p-dichlorobenzene, Halogenated hydrocarbons, such as dibromoethane, dibromobutane, dibromopropane, and dibromobenzene, are illustrated.

From the viewpoint of economy, easy handling, and the like, a hydrocarbon solvent or an ether solvent is preferable. From the viewpoint of reaction rate, 6 to 10 carbon atoms such as benzene, toluene, o-xylene, m-xylene, p-xylene, mesitylene, ethylbenzene, hexane, octane, isooctane, n-nonane and n-decane The hydrocarbon solvent which has is more preferable.

As for the usage-amount of a crosslinkable epoxy compound (B), 0.1-100 mass% is preferable with respect to a granular crosslinked (co) polymer, More preferably, it is 0.5-50 mass%. If the amount is less than 0.1% by mass, the surface crosslinking effect does not appear, and acid / alkali resistance is lowered. If the amount is more than 100% by mass, post-treatment after the reaction becomes very cumbersome.

The amount of the solvent may be used in an amount sufficient to immerse the granular crosslinked polymer in the solvent, for example, two times or more of the mass of the granular crosslinked polymer. From the viewpoint of easy handling, the amount of the solvent used is preferably 3 to 10 times the mass of the granular crosslinked polymer, and more preferably 4 to 7 times.

The concentration of the Lewis acid used as the catalyst may be 0.1 to 100% by mass with respect to the crosslinkable epoxy compound (B), but from the viewpoint of the reaction rate and economical efficiency, the concentration is preferably 1 to 70% by mass. The reaction is preferably carried out at 10 to 100 ° C. for 1 to 16 hours, more preferably at 20 to 60 ° C. for 2 to 10 hours.

Next, the epoxy group of the obtained granular surface crosslinked polymer is hydrolyzed with an acid. Examples of acid catalysts are sulfuric acid, perchloric acid, hydrochloric acid, toluenesulfonic acid and benzenesulfonic acid. Among these, hydrochloric acid is preferable from the viewpoint of easy handling.

As for the density | concentration of the acid used, 0.01-5N are preferable, More preferably, it is 0.05-2N. If the concentration is less than 0.01N, the reaction does not proceed quickly. On the other hand, if the concentration is more than 5N, hydrolysis of the ester bond is likely to occur. At this time, if at least 10 mass% or more of water is contained as a solvent, you may use an organic solvent together.

The organic solvent used is not particularly limited as long as it is compatible with both acid and water and is inert to the acid and oxirane rings. Examples are acetone, methyl ethyl ketone, 1,4-dioxane and tetrahydrofuran.

The granular epoxy ring-opening surface crosslinked (co) polymer thus obtained is reacted with an epoxy compound (C) having 6 to 40 carbon atoms in total to obtain a granular hydrophobic polymer.

Examples of the epoxy compound (C) having 6 to 40 carbon atoms in total include the compounds represented by the following general formulas (1) to (4). From the viewpoint of easy availability, an epoxy compound represented by the general formula (1) or (2) and having 6 to 22 carbon atoms in total is preferable, and a compound having 8 to 18 carbon atoms in total is more preferable.

(Where n represents an integer of 4 to 38)

(Where n represents an integer of 3 to 37.)

(Where n represents an integer of 0 to 32)

(Where n represents an integer of 0 to 31)

The reaction of the epoxy compound and the granular epoxy ring-opening surface crosslinked (co) polymer can be carried out under alkaline conditions or acidic conditions, but from the viewpoint of reaction rate and reproducibility of reaction, it is preferable that the reaction is carried out under acidic conditions. A representative example is the method of carrying out the reaction in the presence of Lewis acid as a catalyst. Examples of Lewis acids used herein include ether complexes of boron trifluoride, tin tetrachloride and titanium tetrachloride. The Lewis acid to be used is not particularly limited, and two or more Lewis acids may be used in combination. In view of easy availability, ether complexes of boron trifluoride are preferred.

Examples of the solvent used in the reaction of the epoxy compound (C) having a total carbon number of 6 to 40 with the granular epoxy ring-opening surface crosslinked polymer include benzene, toluene, o-xylene, m-xylene, p-xylene, mesitylene, Ethylbenzene, propylbenzene, cumene, butylbenzene, isobutylbenzene, amylbenzene, isoamylbenzene, pentane, hexane, cyclohexane, cyclopentane, decalin, heptane, octane, isooctane, n-nonane, n-decane, n Hydrocarbon solvents having 5 to 18 carbon atoms, such as undecane, n-dodecane, n-tridecane, n-tetradecane, n-hexadecane and n-octadecane; Anisole, ethyl isoamyl ether, ethyl-t-butyl ether, diisoamyl ether, diisopropyl ether, diphenyl ether, dibutyl ether, dipropyl ether, dibenzyl ether, tetrahydrofuran, methyl-t- Ether solvents such as butyl ether, 1,4-dioxane, diethylene glycol diethyl ether and triethylene glycol dimethyl; And chloroform, carbon tetrachloride, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, chlorobenzene, o-chlorotoluene, p-chlorotoluene, o-dichlorobenzene, m-dichlorobenzene, p-dichlorobenzene, Halogenated hydrocarbons, such as dibromoethane, dibromobutane, dibromopropane, and dibromobenzene, are illustrated.

From the viewpoint of economy, easy handling, and the like, a hydrocarbon solvent or an ether solvent is preferable. From the viewpoint of reaction rate, 6 to 10 carbon atoms such as benzene, toluene, o-xylene, m-xylene, p-xylene, mesitylene, ethylbenzene, hexane, octane, isooctane, n-nonane and n-decane The hydrocarbon solvent which has is more preferable.

An epoxy compound (C) may be used independently and may be used in combination of 2 or more type among them.

As for the usage-amount of an epoxy compound (C), 10-2,000 mass% is preferable with respect to a crosslinkable epoxy compound (B), More preferably, it is 100-1,000 mass%. If the usage amount is less than 10 mass%, reaction reproducibility is inferior, and if the usage amount exceeds 2,000 mass%, washing | cleaning at the time of a post process becomes cumbersome.

The amount of the solvent to be used may be an amount sufficient to deposit the granular crosslinked polymer in the solvent, and for example, two times or more of the mass of the granular crosslinked polymer. From the viewpoint of easy handling, the amount of the solvent used is preferably 3 to 10 times the mass of the granular crosslinked polymer, and more preferably 4 to 7 times.

The concentration of the Lewis acid used as the catalyst may be 0.1 to 100% by mass with respect to the crosslinkable epoxy compound (B), but from the viewpoint of the reaction rate and economical efficiency, the concentration is preferably 1 to 70% by mass. The reaction is preferably carried out at 10 to 100 ° C. for 1 to 16 hours, more preferably at 20 to 60 ° C. for 2 to 10 hours.

The spherical particles having a particle diameter of 1 to 2,000 µm and preferably 3 to 25 µm thus obtained are classified as necessary and can be used as a filler for reverse phase liquid chromatography. Examples of the eluate include water / acetonitrile, water / methanol, acetonitrile / (aqueous acid or alkaline aqueous solution) and methanol / (aqueous acid or alkaline aqueous solution).

The column filled with the filler of the present invention satisfactorily suppresses swelling / shrinkage depending on the solvent, can be used at pH 1-13, and has excellent acid / alkali resistance. In addition, the ratio of the theoretical number of pyrenes to the theoretical plate number of naphthalene is 0.7 or more, and a sharp peak can be obtained with respect to a polycyclic aromatic compound.

The present invention will be described in more detail below with reference to Examples, but the present invention is not limited to these Examples.

Example 1

<Step 1: Synthesis of Substrate Granular Crosslinked Polymer>

An oil phase was prepared by dissolving 30 g of 2,2'-azobis (isobutyronitrile) in a mixed solution containing 2,000 g of glycerol dimethacrylate and 1,000 g of 1-hexanol. Individually, 120 g of polyvinyl alcohol (Kuraray Poval PVA-224, manufactured by Kuraray) was dissolved in 3 L of water, and 7 L of water was added thereto, followed by addition and mixing of an aqueous solution (2 L) containing 240 g of sodium chloride in an aqueous phase. Was prepared. The oil phase and the water phase thus obtained are mixed in a 20 L volume stainless steel container, and the mixture is mixed in a high speed disperser (homogenizer) while adjusting the rotation speed and dispersion time to obtain an oil droplet having a maximum particle size of 4 µm. Dispersed.

Then, the dispersion was stirred at 150 rpm to proceed the reaction for 7 hours at 60 ℃. The granular crosslinked polymer prepared was centrifuged (at 2,000 rpm for 10 minutes), the supernatant was discarded, the precipitate was dispersed in 12 L of warm water at 70 ° C. (using an ultrasonic cleaner), and the dispersion was stirred at 70 ° C. for 3 hours. It was then filtered by suction. The cake on the funnel was washed with 60 L of hot water at 70 ° C. followed by 18 L of acetone, and the cake was spread out on a stainless steel vat, air dried and further dried at 60 ° C. under reduced pressure for 24 hours. The particles were classified by a wind classifier to obtain 620 g of a granular crosslinked polymer (hereinafter referred to as "substrate gel") having an average particle diameter of 4 µm.

<Step 2: Thorough Cleaning>

500 ml of pure water was added to 50 g of the base gel obtained in Step 1. The resulting mixture was stirred at 60 ° C. under heating for 5 hours, after which the particles were collected by filtration, washed with 2,000 ml of hot water at 70 ° C. and then with 300 ml of methanol, the particles were spread on a stainless steel batt and air dried. Further, 48 g of a thoroughly washed base gel was obtained by drying at 70 ° C. under reduced pressure for 24 hours.

<Step 3: Surface Crosslinking>

In 100 ml of toluene, 20 g of the thoroughly washed base gel obtained in step 2 was dispersed. 2 g of ethylene glycol diglycidyl ether was added thereto while stirring, the mixture was stirred at room temperature for 30 minutes, and a solution obtained by dissolving 500 mg of boron trifluoride diethyl ether complex in 5 ml of toluene was added dropwise thereto. The reaction was carried out for 3 hours at ℃.

Insoluble material was collected by filtration, washed with 100 ml of toluene followed by 100 ml of tetrahydrofuran. The granular polymer was then collected by filtration and transferred to a 300 ml volume beaker. After adding 100 ml of tetrahydrofuran there, the mixture was sonicated for 10 minutes by the ultrasonic cleaner. The granular polymer was collected again by filtration, washed with 100 ml of tetrahydrofuran and then with 100 ml of acetone, air dried, and further dried under reduced pressure at 60 ° C. for 2 hours (amount: 21.2 g).

<Step 4: epoxy ring opening reaction>

15 g of the granular surface crosslinked polymer obtained in Step 3 was dispersed in 60 mL of 0.05 N hydrochloric acid aqueous solution, and stirred at 50 ° C. for 1 hour. The obtained granular epoxy ring-opening polymer was collected by filtration, washed with 500 ml of pure water, air dried, and dried under reduced pressure at 60 ° C. for 2 hours (amount: 15.5 g). Hereinafter, the granular epoxy ring-opening polymer thus obtained is referred to as "unmodified gel".

<Step 5: C18 introduction reaction>

15 g of the non-aqueous gel obtained in step 4 was dispersed in 100 ml of toluene. 105 g of stearyl glycidyl ether was added thereto, and the mixture was stirred at 40 ° C. for 0.5 hour, and thereto was added a solution obtained by dissolving 1 g of boron trifluoride diethyl ether complex in 5 ml of toluene, and at 40 ° C. The reaction was carried out for 5 hours. Thereafter, 100 ml of toluene was added to the reactor, and the granular polymer was collected by filtration, washed with 200 ml of tetrahydrofuran and then transferred to a 300 ml volume beaker. After adding 100 ml of tetrahydrofuran there, the mixture was sonicated for 10 minutes by an ultrasonic cleaner, and the granular polymer was collected again by filtration.

The granular polymer was washed with 100 ml of tetrahydrofuran, transferred again into a 300 ml volume beaker and dispersed in a modified alcohol / 1% aqueous dipotassium hydrogen phosphate solution (50/50 (v / v)). The obtained dispersion was sonicated for 10 minutes by an ultrasonic cleaner, and the granular polymer was collected again by filtration. The collected particulate polymer was washed with 500 ml of pure water and then with 200 ml of acetone, air dried and dried under reduced pressure at 60 ° C. for 2 hours (granular crosslinked polymer with C18 introduced: hereinafter “modified gel gel) "yield: 17.5 g).

<Introduction cost of C18 plane>

From the basic analytical values of the unmodified gel and the modified gel, the introduction ratio of the C18 group was calculated and found to be 20% by mass.

<Filling formula gel>

The modified gel obtained in Step 5 was packed in a stainless steel column of 4.6 mm (inner diameter) x 150 mm (length) by the slurry method to prepare a column for reverse phase liquid chromatography (hereinafter referred to as "column A").

<Performance Measurement 1: Theoretical Fraction Ratio of Naphthalene and Pyrene>

For column A, the theoretical singularity for the peaks of naphthalene and pyrene was measured under the following reverse phase liquid chromatography measurement conditions.

Reverse Phase Liquid Chromatography

Eluent: CH ₃ CN / water = 65/35 (v / v)

Flow rate: 1.00 ml / min

Column temperature: 40 ℃

Detector: UV 254nm

Sample: naphthalene (0.4 mg / ml)

Pyrene (0.3 mg / mL)

Injection volume: 5 μl

As a result, the following values were obtained as the theoretical singularity for each peak. In parenthesis, the ratio when the value of naphthalene is set to 1 is shown. Naphthalene: 7,300 (1.00), pyrene: 6,200 (0.85).

<Performance Measurement 2: Acid Resistance Test>

Acid resistance was tested for column A under the following conditions.

Eluent: MeOH / 1% trifluoroacetic acid aqueous solution (pH 1.2) = 10/90 (v / v)

Flow rate: 1.00 ml / min

Column temperature: 60 ℃

Detector: UV 254nm

Sample: p-hydroxybenzoic acid (0.5 mg / ml)

Injection volume: 10 μl

Exam time: 70 hours

As a result, the following values were obtained as the holding time of p-hydroxybenzoic acid. In parentheses, the ratio when the holding time of p-hydroxybenzoic acid immediately after initiation is 1 is shown. Immediately after the start of the test: 12.6 minutes (1.00), 70 hours after the start of the test: 12.3 minutes (0.98).

<Performance Measurement 3: Alkali Resistance Test>

Alkali resistance was tested for column A under the following conditions, and the retention time of pyrene measured under the same conditions as Performance Measurement 1 before and after the test was compared.

Eluent: CH ₃ CN / 0.1N-NaOH (pH 13) = 20/80 (v / v)

Flow rate: 0.50ml / min

Column temperature: 25 ℃

Exam time: 18 hours

As a result, the following values were obtained as the holding time of pyrene. The values in parentheses indicate the ratio when the value before the test is 1. Before alkali resistance test: 11.6 minutes (1.00), after 18 hours of alkali resistance test: 11.6 minutes (1.00).

<Performance Measurement 4: Solvent Exchange Test>

Solvent exchange was tested for column A under the following conditions, and the theoretical numbers of each of the naphthalene and pyrene measured under the same conditions as Performance Measurement 1 before and after the test were compared. Pure water and methanol were exchanged every 60 minutes at a flow rate of 0.5 ml / min and repeated 5 cycles for each sample.

As a result, the following values were obtained as the theoretical singularity for each peak. In parentheses, the value before the solvent exchange test and the ratio of the stage before the test to the stage after the test are shown. Naphthalene: 7,300 (7,300, 1.00) and pyrene: 6,200 (6,200, 1.00).

Comparative Example 1

<Step 1: Synthesis of Substrate Granular Crosslinked Polymer>

The substrate granular crosslinked polymer (hereinafter, simply referred to as "substrate gel") was obtained by the same operation as in Example 1.

<Step 2: Thorough Cleaning>

The base gel thoroughly washed by the same operation as Example 1 was obtained.

<Step 3: Surface Crosslinking>

20 g of the thoroughly washed base gel obtained in Step 2 was mixed with 20 g of ethylene glycol diglycidyl ether, and the mixture was stirred at room temperature for 30 minutes. Thereafter, 50 g of 1N-NaOH was added thereto and stirred at 30 ° C. for 3 hours.

After collecting by filtration, the insoluble material was washed with 1000 ml of pure water and then with 100 ml of acetone, and dried under reduced pressure at 60 ° C. for 2 hours (amount: 20.7 g).

<Step 4: epoxy ring opening reaction> and

<Step 5: C18 introduction reaction>

By the same operation as in Example 1, a granular crosslinked polymer (hereinafter referred to as "modified gel") into which C18 was introduced was obtained (amount: 18.1 g).

<Introduction cost of C18 plane>

From the basic analytical values of the unmodified gel and the modified gel, it was found that the introduction ratio of the C18 group was 19% by mass.

<Filling formula gel>

The filling was performed by the same operation as in Example 1 (hereinafter referred to as "column B").

<Performance Measurement 1: Theoretical Fraction Ratio of Naphthalene and Pyrene>

Column B was tested under the same conditions as in Example 1.

As a result, the following values were obtained as the theoretical singularity for each peak. In parenthesis, the ratio when the value of naphthalene is set to 1 is shown. Naphthalene: 6,900 (1.00) and pyrene: 5,800 (0.84).

<Performance Measurement 2: Acid Resistance Test>

Column B was tested under the same conditions as in Example 1.

As a result, the following values were obtained as the holding time of p-hydroxybenzoic acid. In parentheses, the ratio when the holding time of p-hydroxybenzoic acid immediately after initiation is 1 is shown. Immediately after the start of the test: 11.4 minutes (1.00), 70 hours after the start of the test: 10.8 minutes (0.95).

<Performance Measurement 3: Alkali Resistance Test>

Column B was tested under the same conditions as in Example 1.

As a result, the following values were obtained as the holding time of pyrene. The values in parentheses indicate the ratio when the value before the test is 1. Before alkali resistance test: 11.0 minutes (1.00), after 18 hours of alkali resistance test: 9.9 minutes (0.90).

<Performance Measurement 4: Solvent Exchange Test>

Column B was tested under the same conditions as in Example 1.

As a result, the following values were obtained as the theoretical singularity for each peak. The values in parentheses indicate the values before the solvent exchange test and the number ratio before and after the test. Naphthalene: 6,500 (6,900, 0.94) and pyrene: 5,000 (5,800, 0.86).

Comparative Example 2

<Step 1: Synthesis of Substrate Granular Crosslinked Polymer>

The substrate granular crosslinked polymer (hereinafter, simply referred to as "substrate gel") was obtained by the same operation as in Example 1.

<Step 2: Thorough Cleaning>

The base gel thoroughly washed by the same operation as Example 1 was obtained.

<Step 3: C18 introduction reaction>

20 g of thoroughly washed base gel obtained in step 2 was dispersed in 110 ml of toluene. After adding 3.9 g of pyridine thereto, the mixture was sonicated for 3 minutes. 6.0 g of stearoyl chloride was dripped there over 15 minutes, stirring, and it was made to react at 60 degreeC for 5 hours. Insoluble material was collected by filtration, sequentially with tetrahydrofuran (250 mL), modified alcohol (250 mL), modified alcohol / water (= 1/1) (250 mL), tetrahydrofuran (250 mL) and methanol It washed with (250 mL) and obtained 35.57 g of modified gel wet with methanol.

<Step 4: Capping>

In 100 ml of 2,2-dimethoxypropane, 35.57 g of a wet modified gel was dispersed with methanol obtained in step 3. After adding 2.0 ml of concentrated hydrochloric acid thereto, the dispersion was sonicated for 3 minutes and then stirred at 50 ° C. for 2 hours. Insoluble material was collected by filtration and washed sequentially with methanol (250 mL), methanol / water (= 1/1) (250 mL) and methanol (250 mL). The gel was then air dried and further dried under reduced pressure at 60 ° C. for 24 hours to give a capped modified gel (21.03 g).

<Introduction cost of octadecanoo diary>

From the respective basic analytical values of the base gel and the modified gel, it was found that the introduction ratio of C18 groups to all hydroxyl groups in the base gel was 14% by mass.

<Filling formula gel>

The filling was performed by the same operation as in Example 1 (hereinafter referred to as "column C").

<Performance measurement 1: theoretical ratio of naphthalene and pyrene>

Column C was tested under the same conditions as in Example 1.

As a result, the following values were obtained as the theoretical singularity for each peak. In parenthesis, the ratio when the value of naphthalene is set to 1 is shown. Naphthalene: 15,100 (1.00) and pyrene: 12,100 (0.80).

<Performance Measurement 2: Acid Resistance Test>

Column C was tested under the same conditions as in Example 1.

As a result, the following values were obtained as the holding time of p-hydroxybenzoic acid. In parentheses, the ratio when the holding time of p-hydroxybenzoic acid immediately after the start of the test is 1 is shown. Immediately after initiation: 11.9 minutes (1.00), 70 hours after the start of the test: 7.1 minutes (0.60).

<Performance Measurement 3: Alkali Resistance Test>

Column C was tested under the same conditions as in Example 1.

As a result, the following values were obtained as the holding time of pyrene. The values in parentheses indicate the ratio when the value before the test is 1. Before alkali resistance test: 11.0 minutes (1.00), after 18 hours of alkali resistance test: 6.4 minutes (0.58).

<Performance Measurement 4: Solvent Exchange Test>

Column C was tested under the same conditions as in Example 1.

As a result, the following values were obtained as the theoretical singularity for each peak. The values in parentheses indicate the values before the solvent exchange test and the number ratio before and after the test. Naphthalene: 11,500 (15,100, 0.76) and pyrene: 6,200 (12,100, 0.51).

Comparative Example 3

Synthesis was carried out in the same manner as in Example 1 except that Step 3 was omitted. As a result, 17.1 g of a modified gel without surface crosslinking treatment was obtained. In step 4, 15 g of the thoroughly washed base gel obtained in step 2 was used as a raw material.

<Introduction cost of C18 plane>

From the basic analytical values of the base gel and the modified gel not subjected to the surface crosslinking treatment, it was found that the introduction ratio of the C18 group was calculated to be 17% by mass.

<Filling formula gel>

The charging was performed by the same operation as in Example 1 (hereinafter referred to as "column D").

<Performance Measurement 1: Theoretical Fraction Ratio of Naphthalene and Pyrene>

Column D was tested under the same conditions as in Example 1.

As a result, the following values were obtained as the theoretical singularity for each peak. In parenthesis, the ratio when the value of naphthalene is set to 1 is shown. Naphthalene: 6,900 (1.00) and pyrene: 5,500 (0.80).

<Performance Measurement 2: Acid Resistance Test>

Column D was tested under the same conditions as in Example 1.

As a result, the following values were obtained as the holding time of p-hydroxybenzoic acid. In parentheses, the ratio when the holding time of p-hydroxybenzoic acid immediately after initiation is 1 is shown. Immediately after the start of the test: 10.6 minutes (1.00), 70 hours after the start of the test: 8.5 minutes (0.80).

<Performance Measurement 3: Alkali Resistance Test>

Column D was tested under the same conditions as in Example 1.

As a result, the following values were obtained as the holding time of pyrene. The values in parentheses indicate the ratio when the value before the test is 1. Before alkali resistance test: 9.5 minutes (1.00), after 18 hours of alkali resistance test: 6.9 minutes (0.69).

<Performance Measurement 4: Solvent Exchange Test>

Column D was tested under the same conditions as in Example 1.

As a result, the following values were obtained as the theoretical singularity for each peak. The values in parentheses indicate the values before the solvent exchange test and the number ratio before and after the test. Naphthalene: 11,500 (15,100, 0.76) and pyrene: 5,100 (6,900, 0.74).

Various evaluation results obtained in Example 1 and Comparative Examples 1 to 3 are shown in Table 1 together.

The ratio of naphthalene and pyrene: pyrene / naphthalene Acid resistance test: retention time ratio of p-hydroxybenzoic acid before and after the test Alkali resistance test: retention time ratio of pyrene before and after test Solvent exchange test: Water ratio before and after the test (top: naphthalene, bottom: pyrene) Column A (Example 1) 0.85 0.98 1.00 0.991.00 Column B (Comparative Example 1) 0.84 0.95 0.90 0.940.86 Column C (Comparative Example 2) 0.80 0.60 0.58 0.760.51 Column D (Comparative Example 3) 0.80 0.80 0.69 0.760.74

In column A obtained in Example 1, the number ratio of naphthalene and pyrene is high, and the holding | maintenance after an acid / alkali tolerance test does not fall. In addition, even if solvent exchange is performed between pure water and methanol, the swelling and shrinkage peculiar to the polymer filler are suppressed and the theoretical number of sheets is not reduced.

It can be seen that the column C filled with the modified gel obtained by performing the surface crosslinking treatment under alkaline conditions has a lower acid / alkali resistance and a higher resistance to solvent exchange than the column A.

According to the manufacturing method of the filler for reverse phase liquid chromatography of this invention, the high performance filler for reverse phase liquid chromatography can be manufactured. The column for reverse phase liquid chromatography filled with the filler for reverse phase liquid chromatography of the present invention can be used in both very low and very high pH ranges.

By using the analytical method by reversed phase liquid chromatography of the present invention, separation / analysis can be performed very precisely, particularly with respect to medicine / pesticides, food additives and their intermediates, natural or synthetic polymers and their additives, and environmental contaminants. . Thus, the present invention is useful in a wide range of fields.

Claims (24)

After reacting the granular crosslinked polymer (A) having a hydroxyl group with the crosslinkable epoxy compound (B) and hydrolyzing the oxirane ring, the compound having the obtained hydroxyl group and the epoxy compound (C) having 6 to 40 carbon atoms in total. A granular hydrophobic polymer prepared by reacting the same. The granular crosslinked polymer (A) having a hydroxyl group according to claim 1, wherein the poly (meth) acrylate (I), glycidyl (meth) acrylate (II) and the molecule of a polyhydric alcohol having at least one hydroxyl group in a molecule thereof. At least two copolymers selected from the group consisting of poly (meth) acrylates (III) of polyhydric alcohols having no hydroxyl group, or homopolymers of poly (meth) acrylates (I) of polyhydric alcohols. Granular hydrophobic polymer. The poly (meth) acrylate (I) of the polyhydric alcohol having one or more hydroxyl groups in the molecule is glycerol dimethacrylate, and the glycidyl (meth) acrylate (II) is glycidyl. Poly (meth) acrylate (III) of the polyhydric alcohol which is methacrylate and does not have a hydroxyl group in the molecule is a granular hydrophobic polymer, characterized in that alkylene glycol dimethacrylate. 4. The particulate hydrophobic polymer according to claim 3, wherein the alkylene glycol dimethacrylate is ethylene glycol dimethacrylate. The granular hydrophobic polymer according to claim 1, wherein the crosslinkable epoxy compound (B) is a compound selected from epihalohydrin and an epoxy compound containing two or more oxirane rings. The epoxy compound according to claim 5, wherein the epoxy compound containing two or more oxirane rings is ethylene glycol diglycidyl ether, propylene glycol diglycidyl ether, butylene glycol diglycidyl ether, trimethylolpropane diglyci. A granular hydrophobic polymer characterized by being dil ether, trimethylolpropane triglycidyl ether, pentaerythritol triglycidyl ether, or triglycidyl isocyanurate. The granular hydrophobic polymer according to claim 1, wherein the epoxy compound (C) having 6 to 40 carbon atoms in total is a compound selected from the group consisting of compounds represented by the following general formulas (1) to (4). . (Where n represents an integer of 4 to 38.) (Where n represents an integer of 3 to 37.) (Where n represents an integer of 0 to 32) (Where n represents an integer of 0 to 31) The granular hydrophobic polymer according to claim 7, wherein the epoxy compound (C) having 6 to 40 carbon atoms in total is stearyl glycidyl ether. The granular hydrophobic polymer according to any one of claims 1 to 8, wherein the average particle diameter is 1 to 2,000 µm. After reacting a granular crosslinked polymer (A) having a hydroxyl group with a crosslinkable epoxy compound (B), hydrolyzing the oxirane ring, and further reacting with an epoxy compound (C) having 6 to 40 carbon atoms in total. A method for producing a granular hydrophobic polymer. The granular crosslinked polymer (A) having a hydroxyl group according to claim 10 is a poly (meth) acrylate (I), glycidyl (meth) acrylate (II) and a molecule of a polyhydric alcohol having one or more hydroxyl groups in a molecule. Granular hydrophobic polymer, characterized in that at least two copolymers selected from poly (meth) acrylate (III) of a polyhydric alcohol having no hydroxyl group, or a homopolymer of poly (meth) acrylate (I) of a polyhydric alcohol. Manufacturing method. The method for producing a granular hydrophobic polymer according to claim 10, wherein the crosslinkable epoxy compound (B) is a compound selected from epihalohydrin and an epoxy compound containing two or more oxirane rings. 12. The poly (meth) acrylate (I) of the polyhydric alcohol having one or more hydroxyl groups in the molecule is glycerol dimethacrylate, and the glycidyl (meth) acrylate (II) is glycidyl. The poly (meth) acrylate (III) of the polyhydric alcohol which is a methacrylate and does not have a hydroxyl group in the said molecule | numerator is a manufacturing method of the granular hydrophobic polymer characterized by the above-mentioned alkylene glycol dimethacrylate. The method for producing a granular hydrophobic polymer according to claim 13, wherein the alkylene glycol dimethacrylate is ethylene glycol dimethacrylate. The method of claim 12, wherein the epoxy compound containing two or more oxirane rings are ethylene glycol diglycidyl ether, propylene glycol diglycidyl ether, butylene glycol diglycidyl ether, trimethylolpropane diglyci Dyl ether, trimethylolpropane triglycidyl ether, pentaerythritol triglycidyl ether, or triglycidyl isocyanurate. The granular hydrophobicity according to claim 10, wherein the epoxy compound (C) having 6 to 40 carbon atoms in total is a compound selected from compounds represented by the general formulas (1) to (4) according to claim 7. Method of Making Polymers. The method for producing a granular hydrophobic polymer according to claim 16, wherein the epoxy compound (C) having 6 to 40 carbon atoms in total is stearyl glycidyl ether. The granular hydrophobic polymer according to claim 10, wherein the reaction of the granular crosslinked polymer (A) having a hydroxyl group with an epoxy compound containing two or more oxirane rings is carried out in the presence of Lewis acid in a low polar solvent. Manufacturing method. The process for producing a particulate hydrophobic polymer according to claim 10, wherein the reaction with the epoxy compound (C) having 6 to 40 carbon atoms in total is carried out in the presence of Lewis acid in a low polar solvent. The method for producing a particulate hydrophobic polymer according to claim 18 or 19, wherein the low polar solvent is a hydrocarbon having 5 to 10 carbon atoms in total. The method for producing a granular hydrophobic polymer according to claim 18 or 19, wherein the concentration of the Lewis acid is 1 to 70% by mass relative to the granular crosslinked polymer. The method for producing a granular hydrophobic polymer according to any one of claims 10 to 21, wherein the granular hydrophobic polymer has an average particle diameter of 1 to 2,000 µm. A column for reverse phase liquid chromatography, wherein the granular hydrophobic polymer according to any one of claims 1 to 9 is filled. As a method of analyzing a sample containing a polycyclic aromatic compound, The analysis method characterized by using the column for reverse phase liquid chromatography of Claim 23.