JP3666023B2 - Chromatographic packing material - Google Patents

Description

（式中、ＲはＲ^a，
【化７】

のいずれかであり、Ｒ^a は置換もしくは非置換アルキル基、置換もしくは非置換フェニル基、及び置換もしくは非置換複素環式基の中から選ばれた置換基である。下記式
【化８】

で表される基は、多孔質担体表面にアミロース誘導体がイソシアナート基を有するシラン処理剤（A1）を介して結合している場合には、該シラン処理剤（A1）由来のケイ素原子と該シラン処理剤（A1）のイソシアナート基由来の−ＮＨＣＯ−基との間の基を意味し、多孔質担体にアミロース誘導体がシラン処理剤（A2）と架橋剤としての多官能イソシアナート誘導体（Ｂ）とを介して結合している場合には、該シラン処理剤（A2）由来のケイ素原子と該多官能イソシアナート誘導体（Ｂ）の１つのイソシアナート基に由来する−ＮＨＣＯ−基との間の基を意味する。前記Ｒ^bは置換もしくは非置換メチレン基、置換もしくは非置換フェニレン基、ヘテロ原子、−ＮＨ−基またはカルボニル基を示す。前記ｍ₂は共有結合しているＲ^bの総数を意味し、１〜２０の整数を示す。ｍ₁はアミロース誘導体の平均重合度を意味し、平均で３〜１０００の整数を示す。Ｚはシラン処理剤（A1）または（A2）由来のケイ素原子に結合している基を意味し、多孔質担体表面の基に由来する基、アルコキシ基、ハロゲン原子、アルキル基、フェニル基、シラン処理剤に由来する基のいずれかを示す）
【０００５】
本発明は、また、アミロースの６位の水酸基の一部あるいは全部を保護基を用いて保護し、残りの水酸基に前記一般式（１）における置換基Ｒ ^a を導入した後、前記保護基をはずし、６位の水酸基の一部で、イソシアナート基を有するシラン処理剤（A1）を介して、またはシラン処理剤（A2）と架橋剤としての多官能イソシアナート誘導体（Ｂ）とを介して多孔質担体の孔内外表面に化学結合させることを特徴とする上記物質の製造法を提供する。
【０００６】
本発明は、さらに、上記物質を有効成分とするクロマト用充填剤を提供する。
【０００７】
なお、上記一般式（１）で表される化合物において、ｍ₁個存在する６位の炭素の少なくとも１個所は、上記式に示したように、シラン処理剤または架橋剤の末端と結合しており、残りの６位の炭素はＯＲであるが、これは略してある。
【０００８】
以下に、本発明の新規物質の１例として、下記の一般式（２）で表される化合物について、その製造法の１例と共に具体的に説明する。
【化９】

（式中、Ｒ¹は
【化１０】

を示す。ｍ ₁ は前記と同じ。Ｅｔはエチル基を示す）
【０００９】
なお、上記一般式（２）で表される化合物において、ｍ₁個存在する６位の炭素の少なくとも１個所は、上記式に示したように、架橋剤末端と結合しており、残りの６位の炭素はＯＲであるが、これは略してある。まず、アミロースの６位の水酸基にトリチル基を導入して保護し、続いて残りの水酸基を修飾剤として３，５−ジメチルフェニルイソシアナートを用いて置換した。次に、得られたアミロース誘導体のトリチル基を酸に浸漬してはずし、表面処理したシリカゲルにコーティングした後、架橋剤としてジフェニルメタン−４，４’−ジイソシアナートを用いてアミロースの６位の水酸基と表面処理シリカゲルを化学結合させることによって目的とする上記一般式（２）で表される新規物質を得た。この場合の反応スキームを以下に示す。なお、一般式（２）で表される化合物において、ｍ₁個存在する６位の炭素の少なくとも１個所は、上記式に示したように、架橋剤末端と結合しており、残りの６位の炭素はＯＲであるが、これは略してある。
【００１０】
【化１１】

【００１１】
この方法により合成した上記一般式（２）で表される本発明の新規化合物は、多孔質シリカゲル孔内外表面に糖部の６位の水酸基の一部で、架橋剤を介して化学結合したものであり、前記の課題を解決したものである。
【００１２】
本発明に用いられるアミロースとは、合成アミロース、天然アミロースのいずれであってもよい。また、アミロース含量の高い澱粉も使用することができる。これらアミロースの数平均重合度としては３以上であり、上限については特に制限はないが、取扱いの点から１０００程度が好ましい。
【００１３】
また、本発明に使用する多孔質担体は、多孔質無機担体や多孔質有機担体であり、具体的にはシリカゲル，けいそう土，多孔質ガラス，ヒドロキシアパタイト，アルミナ，酸化チタン，マグネシア等の多孔質無機担体やポリアクリルアミド、ポリアクリレート等の多孔質有機担体があり、特にシリカゲルが好ましい。シリカゲルとしては、その粒径が１〜１０００μｍ、好ましくは２〜１００μｍであり、平均孔径は１０オングストローム〜１００μｍ、好ましくは５〜５００ｎｍのものが用いられる。なお、多孔質担体として多孔質膜を使用して新規なクロマト用分離膜とすることも可能である。
【００１４】
本発明中のシリカゲルへの表面処理剤としては、アミノ基を持つもの、水酸基をもつもの、メルカプタン基を持つものが好ましいが、従来シランカップリング剤として市販されているものや下記の一般式で示されるものを用いて、架橋剤が結合できるよう合成したものであってもよい。なお、前記一般式中のＲ ^b は、置換もしくは非置換メチレン基、置換もしくは非置換フェニレン基、ヘテロ原子等が共有結合したものを示すが、具体的には下記に示すシラン処理剤の側鎖の一部あるいはスペーサー（架橋剤）とシラン処理剤が化学結合したものの一部を示す。
【００１５】
【化１２】

【００１６】
【化１３】

【００１７】
【化１４】

【００１８】
【化１５】

【００１９】
【化１６】

【００２０】
【化１７】

【００２１】
【化１８】

【００２２】
上記各式中、ｎ₁は１〜３の整数、Ｒ¹は水素原子または炭素数１〜２０程度のアルキル鎖またはその誘導体、Ｒ²は炭素数１〜２０程度のアルキル鎖またはその誘導体、Ｘは炭素数１〜１０の置換もしくは非置換アルコキシ基，ハロゲン原子（好ましくは塩素原子），水酸基，または置換もしくは非置換フェノキシ基（但し、Ｘの少なくとも１つは炭素数１〜１０の置換若しくは非置換アルコキシ基である）、Ｙはハロゲン原子をそれぞれ示す。本発明の新規物質を製造する場合におけるシリカゲルへの表面処理は、公知の方法で行うことができる。
【００２３】
架橋剤としてはアミロースの水酸基と表面処理シリカゲルとを化学結合できるものであればよく、つまり１分子中に２個以上の官能基を持っている化合物であればよいが、本発明では多官能のイソシアナート誘導体を用いる。多官能のイソシアナート誘導体としては、脂肪族あるいは芳香族多官能イソシアナートが挙げられる。例示すれば、ヘキサメチレン−１，６−ジイソシアナート、テトラメチレン−１，４−ジイソシアナート、ジフェニルメタン−４，４’−ジイソシアナート、２，４−トリレンジイソシアナート等のジイソシアナートがある。また、これらジイソシアナートを用いてアミロースと表面処理シリカゲルとを化学結合させる方法は従来公知の方法で行うことができる。
【００２４】
また、架橋剤を使用しないで前記シリカゲルへの表面処理剤として例示した化合物のうち５番目の化合物のようにイソシアナート基を有する処理剤を直接用いてシリカゲルに化学結合させることも可能である。ここで述べる架橋率とは、架橋剤と糖水酸基が１：１で反応すると仮定してアミロース全水酸基数に対する架橋剤の反応率に相当する値である。
【００２５】
上記一般式（１）で表される化合物のアミロース水酸基の一部もしくは全部に導入される置換基（Ｒ ^a ）は該水酸基を修飾するものであり、アミロース部１分子中の水酸基に２種以上の異なる置換基を導入することもできる。該置換基は置換もしくは非置換アルキル基、置換もしくは非置換フェニル基及び置換もしくは非置換複素環式基の中から選ばれ、具体的にはメチル基，エチル基，プロピル基，ブチル基，ｔ−ブチル基，フェニル基，メチルフェニル基，ジメチルフェニル基，エチルフェニル基，ジエチルフェニル基，トリメチルシリルフェニル基，アルコキシフェニル基，ジアルコキシフェニル基，ハロゲン化フェニル基，ジハロゲン化フェニル基，フェニルアゾフェニル基，ナフタレン基，アントラセン基，ピリジル基，フリル基等が挙げられる。上記一般式（１）の糖部水酸基へのこれら置換基の導入法は従来公知の方法で行うことができる。
【００２６】
化学結合するアミロース誘導体の量はシリカゲルに対して１〜５０％、好ましくは５〜３０％である。
【００２７】
【実施例】
以下に実施例を示して本発明を詳しく説明する。
実施例１
一般式（２）で表される新規物質の合成
（１−１）アミロースのトリチル化及びカーバメート誘導体の合成
アミロース（林原社製、商品名：アミロースＢ）３ｇ及びトリチルクロライド１０．３ｇを２００ｍｌ三ッ口ナスフラスコ中で６０℃にて２時間真空乾燥させた後、無水ピリジン６０ｍｌを加えて８０℃で窒素下２９時間反応させた。一部アミロースをサンプリングし、ＩＲスペクトルによりトリチル化されていることを確認し、６−Ｏ−トリチルアミロースを得た。
【００２８】
次に、この反応液に３，５−ジメチルフェニルイソシアナート９．５５ｇを添加し、９０℃で２４時間反応させることにより、残りの水酸基を置換した。一部をサンプリングし、ＩＲスペクトルにより１７１０ｃｍ^-1にカルボニル基の伸縮振動が認められた。この反応液をメタノール１リットル中に落とし込み、沈殿物をメタノールで数回洗浄後、１％ＨＣｌ−メタノール溶液４００ｍｌ中に沈殿物を落とし、室温で１晩攪拌させることによりトリチル基を切断した。得られた沈殿物を中性になるまで洗浄、濾過を繰り返し、真空乾燥して３．８５ｇの沈殿物を得た。¹Ｈ−ＮＭＲ（溶媒：ピリジン−ｄ５、５００ＭＨｚ、基準物質ＴＭＳ、温度８０℃）よりアミロースの２，３位の炭素に結合した３，５−ジメチルフェニルカーバメート由来のメチルプロトン（１．９０−２．００ｐｐｍ）とアミロースの６位の炭素に結合した３，５−ジメチルフェニルカーバメート由来のメチルプロトン（２．２３ｐｐｍ）との面積比より６位の水酸基が５割程度残っていることが確認された。このようにして６位の水酸基が一部未修飾のアミロースカーバメート誘導体を合成した。
【００２９】
（１−２）アミロースカーバメート誘導体のシリカゲルへの固定化（架橋率３％）
予め活性化（１８０℃、２時間真空乾燥）しておいたシリカゲル（ダイソー社製、平均孔径１００ｎｍ、粒径７μｍ）に常法により３−アミノプロピルトリエトキシシランを結合させて得られた表面処理シリカゲル３．３ｇを上記（１−１）で得られたアミロースのカーバメイト誘導体をテトラヒドロフラン（ＴＨＦ）１０ｍｌに溶解した溶液中に分散させ、次にＴＨＦを留去させることにより表面処理シリカゲルにアミロースのカーバメイト誘導体をコーティングした。このアミロースのカーバメイト誘導体をコーティングした表面処理シリカゲルに無水トルエン１０ｍｌ、無水ピリジン２ｍｌ、ジフェニルメチレン−４，４’−ジイソシアナート０．０１５ｇを添加し９０〜１００℃で５時間反応後、さらに残りの水酸基を３，５−ジメチルフェニルイソシアナート０．４６ｇを添加して３０時間反応させることにより置換した。
【００３０】
反応停止後、ガラスフィルターＧ４で濾過し、ＴＨＦ、メタノール、アセトン、ヘキサンで洗浄し、シリカゲルに未結合のアミロース誘導体を除いた。６０℃で２時間の真空乾燥後の重量は３．４５ｇであった。得られた表面処理シリカゲルのＩＲスペクトルより１７１０ｃｍ^-1にカルボニル基の伸縮振動が認められたことにより、シリカゲルへの結合を確認した。これをクロマト用分離剤として使用し、各種ラセミ化合物に対する分離剤としての評価を行った。結果を第１表に示す。なお、第１表中αは分離係数であり、下式により求められる。ｋ₁及びｋ₂は保持容量比であり、それぞれ下式により求められる。
【００３１】
α（分離係数）＝ｋ₂（保持容量比）／ｋ₁（保持容量比）
ｋ₁（保持容量比）
＝［（先に溶出してきた対掌体の保持時間）−デッドタイム］／デッドタイム
ｋ₂（保持容量比）
＝［（後に溶出してきた対掌体の保持時間）−デッドタイム］／デッドタイム
【００３２】
【表１】

【００３３】
応用例
光学分割用カラムの作製と光学分割能
実施例１の（１−２）で得られた物質を０．４６×２５ｃｍのステンレススチール製空カラムにスラリー充填法にて充填した。なお、充填装置はケムコ社製のパッキングシステムを用いた。このカラムを用いて光学分割能を評価した。高速液体クロマトグラフシステムはウオーターズ５１０ポンプ及び４８６ＵＶ検出器等を使用した。また、比較例としてアミローストリス（３，５−ジメチルフェニルカーバメイト）誘導体を表面処理シリカゲルに物理的に吸着した分離剤を用い、これを同条件で分析した結果を第１表に示した（CHEMISTRY LETTERS pp.1857-1860, 1987参照）。表から明らかなように、本発明の物質の光学分割能は、比較例のものと同等であることがわかった。
【００３４】
さらに、本発明により得られた光学分割用カラムの耐溶媒性を調べるために、ＴＨＦ溶液を流速１ｍｌ／ｍｉｎで２時間流した後の光学分割能を測定したところ、変化が認められず、優れた耐溶媒性を有していることが確認できた。なお、分析は溶離液としてヘキサン：ＩＰＡ＝９０：１０を用い、流速０．５ｍｌ／ｍｉｎ、室温で実施した。
【００３５】
【発明の効果】
本発明によれば、水酸基の一部または全部に特定の置換基が導入されたアミロース誘導体を、その６位の水酸基で架橋剤を介して多孔質担体と化学結合させた新規物質を効率よく得ることができる。この物質は光学分割のためのクロマト用充填剤として有用である。[0001]
[Industrial application fields]
The present invention relates to a novel substance, a method for producing the same, and use thereof. Specifically, an amylose derivative having a specific substituent introduced into part or all of a hydroxyl group is bonded to a porous carrier via a crosslinking agent at the hydroxyl group at the 6-position. The present invention relates to a chemically bonded novel substance, a process for producing the same, and a chromatographic separation agent mainly composed of the novel substance.
[0002]
[Background Art and Problems to be Solved by the Invention]
So far, polysaccharides or chromatographic separating agent obtained by supporting by physical methods derivatives thereof such as cellulose or amylose is known to be useful in the optical resolution silica gel. However, this compound has low solvent resistance, and when used in a liquid chromatography method or the like, since the eluent used is limited, the usefulness of the polysaccharide cannot be sufficiently extracted. Further, since the cleaning liquid for cleaning the contaminated column is also limited, there is a problem in that it causes the column deterioration. In addition, a compound in which a polysaccharide derivative is chemically bonded to silica gel in order to overcome the above problems has been known. However, in this case, since the site to be chemically bonded to silica gel cannot be selected, the higher-order structure of the polysaccharide itself is affected, and the useful properties of the polysaccharide are reduced. Furthermore, due to the unspecified binding site, there are disadvantages such as large variations in the quality of the silica gel compound obtained.
[0003]
Thus, as a result of studies to solve this problem, the present inventors have found that the 6-position hydroxyl group of amylose is not so much involved in optical resolution when used as a chromatographic separation agent. Therefore, for example, silica gel is used as a porous carrier, and a compound in which an amylose derivative is chemically bonded to a part of the hydroxyl group at the 6-position via a cross-linking agent on the inner and outer surfaces of the pore is synthesized and used as a chromatographic separation agent for this compound. The present invention was completed by repeatedly examining the usefulness of the above.
[0004]
[Means for Solving the Problems]
That is, the present invention provides a method in which the amylose derivative is part of the hydroxyl group at the 6-position of amylose and has an isocyanate group on the inner and outer surfaces of the porous carrier via the silane treating agent (A1) or the silane treating agent (A2 ) And a polyfunctional isocyanate derivative (B) as a crosslinking agent, and a substance having a structure represented by the following general formula (1) as a main structure.
[Chemical 6]

(Wherein R is R ^a ,
[Chemical 7]

Is any one of, R ^a is a substitution or unsubstituted alkyl group, a substituted or unsubstituted phenyl group, and a substituted or unsubstituted group selected from among unsubstituted heterocyclic group. The following formula [Chemical Formula 8]

In the case where the amylose derivative is bonded to the surface of the porous carrier via the silane treatment agent (A1) having an isocyanate group, the group represented by This means a group between the isocyanate group of the silane treatment agent (A1) and the —NHCO— group derived from the isocyanate group. The amylose derivative is a porous carrier with a silane treatment agent (A2) and a polyfunctional isocyanate derivative (B Between the silicon atom derived from the silane treating agent (A2) and the —NHCO— group derived from one isocyanate group of the polyfunctional isocyanate derivative (B). Means the group of R ^b represents a substituted or unsubstituted methylene group, a substituted or unsubstituted phenylene group, a hetero atom, an —NH— group or a carbonyl group. M ₂ represents the total number of R ^b covalently bonded and represents an integer of 1 to 20. m ₁ denotes an average degree of polymerization of amylose derivative, an integer of 3 to 1000 on average. Z means a group bonded to a silicon atom derived from the silane treating agent (A1) or (A2), a group derived from a group on the surface of the porous carrier, an alkoxy group, a halogen atom, an alkyl group, a phenyl group, a silane Indicates any group derived from the treatment agent)
[0005]
The present invention also part or all of the 6-hydroxyl group of amylose protected using a protecting group, after the introduction of the substituent R ^a in the formula to the remaining hydroxyl group (1), the protecting group Part of the hydroxyl group at the 6-position, via a silane treatment agent (A1) having an isocyanate group, or via a silane treatment agent (A2) and a polyfunctional isocyanate derivative (B) as a crosslinking agent Provided is a method for producing the above substance, which is chemically bonded to the inner and outer surfaces of the pores of a porous carrier.
[0006]
The present invention further provides a chromatographic packing material containing the above substance as an active ingredient .
[0007]
In the compound represented by the general formula (1), at least one of the 6-position carbons present in m ₁ is bonded to the terminal of the silane treating agent or the crosslinking agent as shown in the above formula. The remaining 6th carbon is OR, but this is omitted.
[0008]
Hereinafter, as an example of the novel substance of the present invention, a compound represented by the following general formula (2) will be specifically described together with an example of its production method.
[Chemical 9]

(Where R ¹ is embedded image

The be shown. m ₁ is the same as above. Et represents an ethyl group )
[0009]
In the compound represented by the general formula (2), at least one of the 6-position carbons present in m ₁ is bonded to the end of the crosslinking agent as shown in the above formula, and the remaining 6 The carbon at the position is OR, but this is omitted. First, a trityl group was introduced into the hydroxyl group at the 6-position of amylose for protection, and then the remaining hydroxyl group was substituted with 3,5-dimethylphenyl isocyanate as a modifier. Then, the resulting amylose derivatives trityl group to remove is immersed in acid, after coating the surface-treated silica gel, the 6-position of amylose with a diphenyl-4,4'-diisocyanate as a crosslinking agent The target novel substance represented by the general formula (2) was obtained by chemically bonding the hydroxyl group and the surface-treated silica gel. The reaction scheme in this case is shown below. In the compound represented by the general formula (2), at least one of the 6-position carbons present in m ₁ is bonded to the end of the crosslinking agent as shown in the above formula, and the remaining 6-positions. The carbon of is OR, but this is omitted.
[0010]
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[0011]
The novel compound of the present invention represented by the above general formula (2) synthesized by this method is one in which a part of the hydroxyl group at the 6-position of the sugar moiety is chemically bonded to the inner and outer surfaces of the porous silica gel through a crosslinking agent. It solves the above-mentioned problem.
[0012]
The amylose used in the present invention may be either synthetic amylose or natural amylose. Also, starch with a high amylose content can be used. The number average polymerization degree of these amyloses is 3 or more, and the upper limit is not particularly limited, but is preferably about 1000 from the viewpoint of handling.
[0013]
The porous carrier used in the present invention is a porous inorganic carrier or a porous organic carrier, and specifically, porous materials such as silica gel, diatomaceous earth, porous glass, hydroxyapatite, alumina, titanium oxide, and magnesia. There are porous organic carriers such as porous inorganic carriers and polyacrylamides and polyacrylates , and silica gel is particularly preferable. As the silica gel, those having a particle diameter of 1 to 1000 μm, preferably 2 to 100 μm and an average pore diameter of 10 Å to 100 μm, preferably 5 to 500 nm are used. In addition, it is also possible to use a porous membrane as a porous carrier to obtain a novel separation membrane for chromatography.
[0014]
As the surface treating agent for the silica gel in the present invention, those having an amino group, those having a hydroxyl group, those having a mercaptan group are preferable, but those conventionally marketed as silane coupling agents and the following general formula What is shown may be synthesized so that a crosslinking agent can be bonded. In the general formula, R ^b represents a group in which a substituted or unsubstituted methylene group, a substituted or unsubstituted phenylene group, a hetero atom or the like is covalently bonded. Specifically, the side chain of the silane treating agent shown below Or a part of a spacer (crosslinking agent) and a silane treatment agent chemically bonded.
[0015]
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[0016]
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[0017]
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[0018]
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[0019]
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[0020]
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[0021]
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[0022]
In the above formulas, n ₁ is an integer of 1 to 3, R ¹ is an alkyl chain or a derivative thereof of about 1-20 hydrogen atoms or carbon atoms, R ² is an alkyl chain or a derivative thereof of about 1 to 20 carbon atoms, X a substituted or unsubstituted alkoxy group having a carbon number of 1-10, halogen atom (preferably chlorine atom), a hydroxyl group or a substituted or unsubstituted phenoxy group (provided that at least one is a substituted or unsubstituted 1 to 10 carbon atoms X, Y is a substituted alkoxy group, and Y represents a halogen atom. The surface treatment on silica gel in the production of the novel substance of the present invention can be performed by a known method.
[0023]
Any crosslinking agent may be used as long as it can chemically bond the hydroxyl group of amylose and the surface-treated silica gel, that is, any compound having two or more functional groups in one molecule . Isocyanate derivatives are used. Examples of the polyfunctional isocyanate derivative include aliphatic or aromatic polyfunctional isocyanates. For example, diisocyanates such as hexamethylene-1,6-diisocyanate, tetramethylene-1,4-diisocyanate, diphenylmethane-4,4′-diisocyanate, 2,4-tolylene diisocyanate, etc. There is. A method of chemically bonding amylose and surface-treated silica gel using these diisocyanates can be performed by a conventionally known method.
[0024]
It is also possible to chemically bond to the silica gel with the treatment agent directly with isocyanate groups as the fifth compound of the compounds exemplified as the surface treatment agent to the silica gel without using a crosslinking agent. The crosslinking rate described here is a value corresponding to the reaction rate of the crosslinking agent with respect to the total number of amylose hydroxyl groups, assuming that the crosslinking agent and sugar hydroxyl groups react 1: 1.
[0025]
The substituent ( R ^a ) introduced into part or all of the amylose hydroxyl group of the compound represented by the general formula (1) modifies the hydroxyl group, and two or more kinds of hydroxyl groups in one molecule of the amylose part are used. It is also possible to introduce different substituents. The substituent is selected from a substituted or unsubstituted alkyl group, a substituted or unsubstituted phenyl group and a substituted or unsubstituted heterocyclic group, and specifically includes a methyl group, an ethyl group, a propyl group, a butyl group, t- Butyl group, phenyl group, methylphenyl group, dimethylphenyl group, ethylphenyl group, diethylphenyl group, trimethylsilylphenyl group, alkoxyphenyl group, dialkoxyphenyl group, halogenated phenyl group, dihalogenated phenyl group, phenylazophenyl group, Naphthalene group, anthracene group, pyridyl group, furyl group and the like can be mentioned. The method for introducing these substituents into the sugar moiety hydroxyl group of the general formula (1) can be carried out by a conventionally known method.
[0026]
The amount of the amylose derivative chemically bound is 1 to 50%, preferably 5 to 30% with respect to the silica gel.
[0027]
【Example】
Hereinafter, the present invention will be described in detail with reference to examples.
Example 1
Synthesis of novel substance represented by general formula (2) (1-1) Tritylation of amylose and synthesis of carbamate derivative 3 g of amylose (trade name: Amylose B, manufactured by Hayashibara) and 10.3 g of trityl chloride After vacuum drying at 60 ° C. for 2 hours in an open mouth flask, 60 ml of anhydrous pyridine was added and reacted at 80 ° C. under nitrogen for 29 hours. A part of amylose was sampled and confirmed to be tritylated by IR spectrum to obtain 6-O-trityl amylose.
[0028]
Next, 9.55 g of 3,5-dimethylphenyl isocyanate was added to this reaction solution and reacted at 90 ° C. for 24 hours to replace the remaining hydroxyl groups. A part was sampled, and a stretching vibration of the carbonyl group was observed at 1710 cm ⁻¹ by IR spectrum. The reaction solution was dropped into 1 liter of methanol, and the precipitate was washed several times with methanol. Then, the precipitate was dropped into 400 ml of 1% HCl-methanol solution and stirred at room temperature overnight to cleave the trityl group. The obtained precipitate was repeatedly washed and filtered until neutral, and vacuum dried to obtain 3.85 g of precipitate. ^{From 1} H-NMR (solvent: pyridine-d5, 500 MHz, reference substance TMS, temperature 80 ° C.), methyl proton (1.90-2) derived from 3,5-dimethylphenylcarbamate bonded to the carbons at the 2nd and 3rd positions of amylose. .00 ppm) and the area ratio of 3,5-dimethylphenylcarbamate-derived methyl proton (2.23 ppm) bonded to the 6-position carbon of amylose, it was confirmed that about 50% of the 6-position hydroxyl group remained. . In this way, an amylose carbamate derivative in which the hydroxyl group at the 6-position was partially unmodified was synthesized.
[0029]
(1-2) Immobilization of amylose carbamate derivative on silica gel (crosslinking rate 3%)
Surface treatment obtained by binding 3-aminopropyltriethoxysilane to silica gel (Daiso Corp., average pore size 100 nm, particle size 7 μm) activated in advance (180 ° C., 2 hours under vacuum) by a conventional method. Disperse 3.3 g of silica gel in a solution of the carbamate derivative of amylose obtained in (1-1) above in 10 ml of tetrahydrofuran (THF), and then distill THF to distill off the amylose carbamate on the surface-treated silica gel. The derivative was coated. 10 ml of anhydrous toluene, 2 ml of anhydrous pyridine and 0.015 g of diphenylmethylene-4,4′-diisocyanate were added to the surface-treated silica gel coated with this amylose carbamate derivative and reacted at 90-100 ° C. for 5 hours. The hydroxyl group was replaced by adding 0.46 g of 3,5-dimethylphenyl isocyanate and reacting for 30 hours.
[0030]
After stopping the reaction, the mixture was filtered with a glass filter G4, washed with THF, methanol, acetone, and hexane to remove amylose derivatives not bound to silica gel. The weight after vacuum drying at 60 ° C. for 2 hours was 3.45 g. From the IR spectrum of the obtained surface-treated silica gel, the binding to the silica gel was confirmed by confirming the stretching vibration of the carbonyl group at 1710 cm ⁻¹ . This was used as a chromatographic separating agent and evaluated as a separating agent for various racemic compounds. The results are shown in Table 1. In Table 1, α is a separation factor, and is obtained by the following equation. k ₁ and k ₂ are retention capacity ratios, which are respectively obtained by the following equations.
[0031]
α (separation factor) = k ₂ (retention capacity ratio) / k ₁ (retention capacity ratio)
k ₁ (Retention capacity ratio)
= [(Retention time of enantiomers eluted earlier) −Dead time] / Dead time k ₂ (Retention capacity ratio)
= [(Retention time of enantiomers eluted later) −Dead time] / Dead time
[Table 1]

[0033]
It was filled with slurry packing method to a stainless steel empty column resulting material 0.46 × 25 cm in applications for optical resolution column manufactured and optical resolution ability of Example 1 (1-2). The packing device used was a packing system manufactured by Chemco. The optical resolution was evaluated using this column. The high performance liquid chromatograph system used a Waters 510 pump and a 486 UV detector. Further, as a comparative example, a separation agent in which an amylose tris (3,5-dimethylphenylcarbamate) derivative was physically adsorbed on a surface-treated silica gel was used. pp.1857-1860, 1987). As is apparent from the table, the optical resolution of the substance of the present invention was found to be equivalent to that of the comparative example.
[0034]
Furthermore, in order to investigate the solvent resistance of the optical resolution column obtained by the present invention, the optical resolution after flowing a THF solution at a flow rate of 1 ml / min for 2 hours was measured. It was confirmed that the product has solvent resistance. The analysis was performed using hexane: IPA = 90: 10 as an eluent and a flow rate of 0.5 ml / min at room temperature.
[0035]
【The invention's effect】
According to the present invention, a novel substance in which an amylose derivative having a specific substituent introduced into a part or all of hydroxyl groups is chemically bonded to a porous carrier at the 6-position hydroxyl group via a crosslinking agent is efficiently obtained. be able to. This material is useful as a chromatographic filler for optical resolution.

Claims (5)

On the inner and outer surfaces of the porous carrier, the amylose derivative is part of the hydroxyl group at the 6-position of amylose, via the silane treatment agent (A1) having an isocyanate group, or as a crosslinking agent with the silane treatment agent (A2). A substance that is chemically bonded via a polyfunctional isocyanate derivative (B) and has a structure represented by the following general formula (1) as a main structure.

(Wherein R is R ^a ,

Is any one of, R ^a is a substitution or unsubstituted alkyl group, a substituted or unsubstituted phenyl group, and a substituted or unsubstituted group selected from among unsubstituted heterocyclic group. Following formula

In the case where the amylose derivative is bonded to the surface of the porous carrier via the silane treatment agent (A1) having an isocyanate group, the group represented by This means a group between the isocyanate group of the silane treatment agent (A1) and the —NHCO— group derived from the isocyanate group. The amylose derivative is a porous carrier with a silane treatment agent (A2) and a polyfunctional isocyanate derivative (B Between the silicon atom derived from the silane treating agent (A2) and the —NHCO— group derived from one isocyanate group of the polyfunctional isocyanate derivative (B). Means the group of R ^b represents a substituted or unsubstituted methylene group, a substituted or unsubstituted phenylene group, a hetero atom, an —NH— group or a carbonyl group. M ₂ represents the total number of R ^b covalently bonded and represents an integer of 1 to 20. m ₁ denotes an average degree of polymerization of amylose derivative, an integer of 3 to 1000 on average. Z means a group bonded to a silicon atom derived from the silane treating agent (A1) or (A2), a group derived from a group on the surface of the porous carrier, an alkoxy group, a halogen atom, an alkyl group, a phenyl group, a silane Indicates any group derived from the treatment agent)   The substance according to claim 1, wherein the porous carrier is silica gel. Following formula (2)

(Where R ¹ is

Indicates. m ₁ denotes an average degree of polymerization of amylose derivative, an integer of 3 to 1000 on average. Et represents an ethyl group)
The substance according to claim 1, which is a substance represented by: Some or all of the 6-hydroxyl group of amylose protected using a protecting group, after the introduction of the substituent R ^a in the general formula of claim 1, wherein the remaining hydroxyl groups (1), remove the protecting group, Part of the hydroxyl group at the 6-position, porous via a silane treating agent (A1) having an isocyanate group, or via a silane treating agent (A2) and a polyfunctional isocyanate derivative (B) as a crosslinking agent 2. The method for producing a substance according to claim 1, wherein the substance is chemically bonded to the inside and outside surfaces of the pores of the carrier.   A chromatographic filler comprising the substance according to any one of claims 1 to 3 as an active ingredient.