JP6218574B2 - Liquid chromatography packing - Google Patents
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- JP6218574B2 JP6218574B2 JP2013245185A JP2013245185A JP6218574B2 JP 6218574 B2 JP6218574 B2 JP 6218574B2 JP 2013245185 A JP2013245185 A JP 2013245185A JP 2013245185 A JP2013245185 A JP 2013245185A JP 6218574 B2 JP6218574 B2 JP 6218574B2
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Description
本発明は液体クロマトグラフィ用充填剤に関する。 The present invention relates to a packing material for liquid chromatography.
液体クロマトグラフィ装置のカラムに充填する充填剤としては一般にシリカ粒子やアルミナ粒子が用いられている。 In general, silica particles and alumina particles are used as the packing material to be packed in the column of the liquid chromatography apparatus.
シリカ粒子やアルミナ粒子を充填剤とした場合には、使用できる移動相としてアルカリ性のものを使用するとシリカやアルミナが溶出してしまうため、使用できる移動相のｐＨ範囲が狭く、また吸着能力も低い。
このため、特許文献１には、スチレン-ジビニルベンゼン共重合体を液体クロマトグラフィの充填剤として用いることが提案されている。
When silica particles or alumina particles are used as fillers, if alkaline substances are used as usable mobile phases, silica and alumina are eluted, so the pH range of usable mobile phases is narrow and the adsorption capacity is low. .
For this reason, Patent Document 1 proposes to use a styrene-divinylbenzene copolymer as a filler for liquid chromatography.
また、特許文献２には、シリカ系充填剤として、シリカコア粒子の表面に多孔質のシリカシェルを形成したものが提案されている。このように表面を多孔質とすることで、送液抵抗を大きくしないで、つまり充填剤の粒子径を小さくしなくとも充填剤粒子の比表面積を大きくすることができるので、吸着能力を大きくできる。 Patent Document 2 proposes a silica-based filler in which a porous silica shell is formed on the surface of silica core particles. By making the surface porous in this way, the specific surface area of the filler particles can be increased without increasing the liquid feeding resistance, that is, without reducing the particle diameter of the filler, so that the adsorption capacity can be increased. .
尚、最近ではシリカ粒子の表面にＯＤＳ（オクタデシルシラン）を化学的に結合させて連続的した多検体処理を可能としている。 Recently, ODS (octadecylsilane) is chemically bonded to the surface of silica particles to enable continuous multi-analyte processing.
また、特許文献３には特許文献２と同様に、コアとシェルをスチレン−ジビニルベンゼン共重合体によって構成した液体クロマトグラフィ用充填剤が提案されている。特許文献３ににあっては、試料成分がコアに浸透しにくくシェルに吸着されやすくするため、コアの架橋度を５０％、シェルの架橋度を２０％に設定している。 Patent Document 3 proposes a liquid chromatography packing material in which a core and a shell are composed of a styrene-divinylbenzene copolymer, as in Patent Document 2. In Patent Document 3, in order to prevent sample components from penetrating into the core and easily adsorbed to the shell, the cross-linking degree of the core is set to 50% and the cross-linking degree of the shell is set to 20%.
また、特許文献３には、スチレン−ジビニルベンゼン共重合体以外の材料として、ポリメタクリレート、ポリビニルアルコール、ポリヒドロキシメタアクリレート、ポリアクリレート、ポリビニルアルコール、ポリエーテル、（メタ）アクリレート系、ビニルアルコール系、アクリルアミド系、メタクリル酸系、アクリル酸系が使用できると記載されている。 Patent Document 3 discloses materials other than styrene-divinylbenzene copolymer, such as polymethacrylate, polyvinyl alcohol, polyhydroxymethacrylate, polyacrylate, polyvinyl alcohol, polyether, (meth) acrylate, vinyl alcohol, It is described that acrylamide, methacrylic acid, and acrylic acid can be used.
液体クロマトグラフィ用カラムの理論段高さ、送液性及び浸透性を向上して、分離精度を高め分離時間を短くするため、特許文献２では、シリカコアの表面に多孔性のシリカシェルを形成している。 In Patent Document 2, a porous silica shell is formed on the surface of the silica core in order to improve the theoretical plate height, liquid feeding property and permeability of the liquid chromatography column, improve the separation accuracy and shorten the separation time. Yes.
尚、特許文献３では、単味のポリマーコアとシェルの架橋度を変えることで内側のコアを密に、外側のシェルを粗にすると記載されているが、架橋度を低くすることと多孔性とは関連性はない。ジビニルベンゼン（ＤＶＢ）の割合を多くすると架橋度が大きくなり樹脂内の分子の網目（ミクロポア）は小さくなり、少なくすると架橋度は小さくなり樹脂内の分子の網目（ミクロポア）は大きくなる。一方、多孔質と称される吸着能を発揮する細孔（マクロポア）は上記網目（ミクロポア）とは異なる。 In Patent Document 3, it is described that the inner core is made dense and the outer shell is made rough by changing the degree of crosslinking between the simple polymer core and the shell. Is not related to Increasing the proportion of divinylbenzene (DVB) increases the degree of crosslinking and decreases the molecular network (micropores) in the resin, while decreasing the ratio decreases the degree of crosslinking and increases the molecular network (micropores) in the resin. On the other hand, pores (macropores) that exhibit adsorption ability called porous are different from the meshes (micropores).
このように従来の液体クロマトグラフィ用充填剤にあっては、充填剤の材料、粒径及びコア表面に多孔質シェルを形成した構造が提案されているが、コアとシェルの材料を異ならせたものは提案されていない。
コアとシェルの材料が同じだと境界面において剥離が起きやすく機械的強度に劣り、充填剤の寿命が短くなる。
As described above, in the conventional liquid chromatography packing material, a structure in which a porous shell is formed on the material, particle size, and core surface of the packing material has been proposed, but the core and shell materials are different. Has not been proposed.
If the core and shell materials are the same, delamination is likely to occur at the interface, resulting in poor mechanical strength and shortened filler life.
液体クロマトグラフィ用充填剤をコアとシェルの２層構造としたのは、夫々に要求される機能が異なるからである。即ちシェルには試料分子を吸着したり、イオン交換する能力が要求され、逆にコアの試料分子吸着能力を高めると、却って分離時間が長くなるので、吸着能力は低い方がよい。
しかしながら、コアとシェルを同じ材料で構成すると上記した機械的な強度に問題が生じ、コア及びシェルに要求される機能の発揮が中途半端になる。
The reason why the liquid chromatography packing material has a two-layer structure of a core and a shell is that the required functions are different. That is, the shell is required to have the ability to adsorb sample molecules or exchange ions. Conversely, if the core's ability to adsorb sample molecules is increased, the separation time becomes longer.
However, if the core and the shell are made of the same material, a problem arises in the mechanical strength described above, and the functions required for the core and the shell are halfway.
上記課題を解決するため本発明に係る液体クロマトグラフィ用充填剤は、無孔質コアの表面に多孔質シェルを形成し、前記無孔質コアはエチレングリコールジメタクリラート、ブチレングリコールジメタクリラート、ヘキサンジオールジメタクリラート又はトリメチロールプロパントリメタクリラートの何れかを重合させたポリマーで、前記多孔質シェルはスチレン−ジビニルベンゼン共重合体からなる。 In order to solve the above problems, the packing material for liquid chromatography according to the present invention forms a porous shell on the surface of a non-porous core, and the non-porous core includes ethylene glycol dimethacrylate, butylene glycol dimethacrylate, hexane. A polymer obtained by polymerizing either diol dimethacrylate or trimethylolpropane trimethacrylate, and the porous shell is made of a styrene-divinylbenzene copolymer.
本明細書において、無孔質とは試料分子（例えば分子量７０〜１００００）が実質的に浸透しないことを指し、多孔質とは試料分子（例えば分子量７０〜１００００）が容易に浸透できることを指す。 In the present specification, nonporous means that a sample molecule (for example, molecular weight 70 to 10000) does not substantially penetrate, and porous means that a sample molecule (for example, molecular weight 70 to 10,000) can easily penetrate.
また、前記多孔質シェルに常法によって、スルホン基などの陽イオン交換基やアミノ基などの陰イオン交換基を導入してもよい。このようにすることで、更に分離速度を高めることができる。 Further, a cation exchange group such as a sulfone group or an anion exchange group such as an amino group may be introduced into the porous shell by a conventional method. In this way, the separation speed can be further increased.
本発明に係る液体クロマトグラフィ用充填剤によれば、外表面を多孔質シェルとしたことで、試料に対する吸着能力が向上しただけでなく、充填剤を構成するコアとシェルの材質を異ならせたことで、コアとシェルの接着強度が高まり且つコアに要求される機能とシェルに要求される機能を十分に発揮することができる。 According to the packing material for liquid chromatography according to the present invention, the outer surface is made of a porous shell, so that not only the adsorption capacity for the sample is improved, but also the materials of the core and the shell constituting the packing material are made different. Thus, the adhesive strength between the core and the shell is increased, and the function required for the core and the function required for the shell can be sufficiently exhibited.
特に、コアの材料をエチレングリコールジメタクリレート、ブチレングリコールジメタクリレート、ヘキサンジオールジメタクリラート又はトリメチロールプロパントリメタクリレートの何れかを重合させたポリマーとしたことで、耐酸性、耐アルカリ性に優れ、機械的な強度に優れた充填剤が得られ、またシェルの材料をスチレン−ジビニルベンゼン共重合体としたことで、試料分子の吸着（分離）機能に優れた充填剤が得られ、更にコアとシェルの境界面においてスチレン−ジビニルベンゼンのモノマーがコアに食い込み（楔効果）、接着強度が高まる。 In particular, the core material is a polymer obtained by polymerizing one of ethylene glycol dimethacrylate, butylene glycol dimethacrylate, hexanediol dimethacrylate, or trimethylolpropane trimethacrylate, so it has excellent acid resistance and alkali resistance, and is mechanical. By using a styrene-divinylbenzene copolymer as the shell material, a filler excellent in the adsorption (separation) function of the sample molecules can be obtained. At the interface, the styrene-divinylbenzene monomer bites into the core (wedge effect), increasing the adhesive strength.
図１及び図２に示すように本発明に係る充填剤はコア１とシェル２からなる直径５μｍ程度の球状粒子である。 As shown in FIG. 1 and FIG. 2, the filler according to the present invention is a spherical particle having a diameter of about 5 μm composed of a core 1 and a shell 2.
コア１はモノマーと重合開始剤を、アニオン性分散剤を溶解した水溶液中に混合し、温度・時間などの条件を適切な範囲にして重合せしめることで得る。モノマーとしてはエチレングリコールジメタクリレート、ブチレングリコールジメタクリレート、ヘキサンジオールジメタクリラート又はトリメチロールプロパントリメタクリレートの何れかを選択する。 The core 1 is obtained by mixing a monomer and a polymerization initiator in an aqueous solution in which an anionic dispersant is dissolved, and polymerizing the mixture under conditions such as temperature and time. As the monomer, ethylene glycol dimethacrylate, butylene glycol dimethacrylate, hexanediol dimethacrylate, or trimethylolpropane trimethacrylate is selected.
シェル２はスチレン−ジビニルベンゼン共重合体からなる。このスチレン−ジビニルベンゼン共重合体は、トルエンをベースとしこれに沈殿剤、膨潤剤および重合に関与しないポリマーを添加して希釈剤を調整し、この希釈剤にスチレン−ジビニルベンゼンのモノマーを均一に溶解し、重合開始剤とともに前記コア粒子を含むアニオン性分散在水溶液中に混合し、温度・時間などの条件を適切な範囲にして重合せしめ、コア表面にスチレン−ジビニルベンゼン共重合体からなる多孔質皮膜（シェル）を形成する。 The shell 2 is made of a styrene-divinylbenzene copolymer. This styrene-divinylbenzene copolymer is based on toluene, and a diluent is prepared by adding a precipitating agent, a swelling agent and a polymer that does not participate in polymerization, and the styrene-divinylbenzene monomer is uniformly added to the diluent. Dissolved and mixed in an aqueous dispersion containing an anionic dispersion containing the core particles together with a polymerization initiator, polymerized under appropriate conditions such as temperature and time, and the core surface is made of a styrene-divinylbenzene copolymer. A pellicle (shell) is formed.
そして必要に応じて、シェル２の細孔表面に、スルホン基などの陽イオン交換基やアミノ基などの陰イオン交換基を導入する。 If necessary, a cation exchange group such as a sulfone group or an anion exchange group such as an amino group is introduced into the pore surface of the shell 2.
（実施例１）
精製水４０００ｍｌを４０℃に保ちながら、Ｎ２ガスを３００ｍｌ／ｍｉｎの流量で３０分水中での脱気を行った後、Ｎ２ガスの吹き出し口を水面より上に上げ２００ｍｌ／ｍｉｎの流量に落とし、分散剤としてドデシルベンゼンスルホン酸ソーダ１０ｇを加え溶解させた後、エチレングリコールジメタクリレート５０ｇに重合開始剤２,２’−アゾビス（２,４-ジメチルバレロニトリル）０．５ｇを加えた。
Example 1
While maintaining purified water 4000ml to 40 ° C., the N 2 gas after degassing at 30 in diversion at a flow rate of 300 ml / min, the flow rate of raising the outlet of the N 2 gas above the surface of the water 200 ml / min After dropping and dissolving 10 g of sodium dodecylbenzenesulfonate as a dispersing agent, 50 g of ethylene glycol dimethacrylate was added with 0.5 g of a polymerization initiator 2,2′-azobis (2,4-dimethylvaleronitrile).
この後、攪拌を行った。攪拌は長さ１２ｃｍ巾２ｃｍの水平状の攪拌翼を用い、１１００ｒｐｍの回転速度で４０℃を保ちながら６０分間重合を行った。 This was followed by stirring. The stirring was performed using a horizontal stirring blade having a length of 12 cm and a width of 2 cm, and polymerization was carried out for 60 minutes while maintaining 40 ° C. at a rotation speed of 1100 rpm.
次いで、５０〜６０％架橋に調整したスチレン−ジビニルベンゼンのモノマーに希釈剤（１０％ポリスチレンを溶解したトルエン）５０ｇに重合開始剤２,２’−アゾビス（２,４-ジメチルバレロニトリル）０．５ｇを溶解し、これを１０００ｒｐｍに回転速度を落とし、２０ｍｌ／ｍｉｎ程度の滴下速度で加え、４０℃を保ちながら６０分間重合し、その後６０℃まで昇温し、４時間重合することで重合を完了した。 Subsequently, a polymerization initiator 2,2′-azobis (2,4-dimethylvaleronitrile) was added to 50 g of a diluent (toluene in which 10% polystyrene was dissolved) in a styrene-divinylbenzene monomer adjusted to 50 to 60% crosslinking. Dissolve 5 g, lower the rotational speed to 1000 rpm, add at a dropping speed of about 20 ml / min, polymerize for 60 minutes while maintaining 40 ° C., then raise the temperature to 60 ° C. and polymerize for 4 hours to polymerize. Completed.
重合物は濾別し水洗後、２-ブタノンで希釈剤や不純物を除去するためカラム洗浄し、この後分級機を用い、粒子径を約５μｍに整えた。
糖分離用充填剤とするため、５μｍ樹脂は完全に脱水し、樹脂量と同量のクロロメチルエーテルを用い、０〜５℃の温度で３時間反応させてクロロメチル化した。
The polymer was separated by filtration, washed with water, washed with column to remove diluent and impurities with 2-butanone, and then the particle size was adjusted to about 5 μm using a classifier.
In order to obtain a sugar separation filler, the 5 μm resin was completely dehydrated, and chloromethylated by reacting at a temperature of 0 to 5 ° C. for 3 hours using the same amount of chloromethyl ether as that of the resin.
クロロメチル化樹脂は十分水洗後脱水し、２０％テトラメチレンジアミノヘキサン水溶液中に移し、６０℃で３時間緩やかに攪拌しながらアミノ化を行った。アミノ化終了後、十分な水洗を行い、５０〜６０％の保水状態で保存する。 The chloromethylated resin was thoroughly washed with water, dehydrated, transferred to a 20% tetramethylenediaminohexane aqueous solution, and aminated with gentle stirring at 60 ° C. for 3 hours. After the amination is completed, the product is sufficiently washed with water and stored in a water retention state of 50 to 60%.
図３は、実施例１で作製したイオン交換基としてアミノ基を導入した本発明に係る充填剤と従来の充填剤との分離能力を比較したグラフである。
従来の充填剤としては、コアとシェルの区別がなく全体が多孔質状のスチレン−ジビニルベンゼン共重合体からなる充填剤を用いた。
FIG. 3 is a graph comparing the separation ability of a filler according to the present invention in which an amino group is introduced as an ion exchange group prepared in Example 1 and a conventional filler.
As a conventional filler, a filler composed of a styrene-divinylbenzene copolymer having a porous structure without distinction between a core and a shell was used.
図３から明らかなように、本発明に係る充填剤は従来の充填剤に比較して、大幅に分離時間が短縮し且つ分離精度は低下していないことが分かる。 As is clear from FIG. 3, it can be seen that the filler according to the present invention significantly reduces the separation time and does not lower the separation accuracy as compared with the conventional filler.
（実施例２）
条件は（実施例１）と同様とした。但し、イオン交換基（アミノ基）を多孔質シェルに導入しない充填剤を作製した。
(Example 2)
The conditions were the same as in (Example 1). However, the filler which does not introduce | transduce an ion exchange group (amino group) into a porous shell was produced.
図４は、実施例２で作製したイオン交換基を導入していない本発明に係る充填剤と従来の充填剤との分離能力を比較したグラフである。
図４から明らかなように、本発明に係る充填剤であってイオン交換基を導入していない充填剤は、実施例１の充填剤よりも分離時間は長くなっているが、従来の充填剤に比較して、大幅に分離時間が短縮し且つ分離精度は低下していないことが分かる。
FIG. 4 is a graph comparing the separation ability of the filler according to the present invention in which the ion exchange group prepared in Example 2 is not introduced and the conventional filler.
As is clear from FIG. 4, the filler according to the present invention, which is not introduced with an ion exchange group, has a longer separation time than the filler of Example 1, but the conventional filler. It can be seen that the separation time is significantly shortened and the separation accuracy is not lowered as compared with FIG.
本発明に係る液体クロマトグラフィ用充填剤は、分離能を損なうことなくボイド時間を短縮できるため、高速液体クロマトグラフィに好適に適用できる。 The packing material for liquid chromatography according to the present invention can be suitably applied to high performance liquid chromatography because the void time can be shortened without impairing the resolution.
1…コア、２…シェル。 1 ... core, 2 ... shell.
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