JP3048085B2 - Method for immobilizing cyclodextrin - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a method for immobilizing cyclodextrin, and more particularly, to a method for immobilizing cyclodextrin on an inorganic substance, particularly silica gel.

[0002]

2. Description of the Related Art Cyclodesquitrin incorporates various organic compounds into its annular cavity to form an inclusion compound,
These compounds have recently been used as adsorption / separation agents. However, since cyclodextrin itself is water-soluble and is dissolved only in a specific organic solvent, it is difficult to include and isolate a compound from various systems.

As one method for solving such a problem, a method has been proposed in which cyclodextrin is chemically bonded to an inorganic substance such as silica gel to make it insoluble and easily used as an adsorbing / separating agent. I have.

A method for chemically bonding cyclodextrin to silica gel is disclosed in, for example, US Pat. No. 4,539,399.
Specification, JP-A-61-129566, and Journal Oblique Liquid Chromatography (J. Liquid chromato
gr., 9 (2 & 3) 607-620 (1986)), 3-glycidoxypropyltrimethoxysilane, γ-aminopropyltriethoxysilane, 3-isocyanatopropyltriethoxysilane as a silylating agent for silica gel. And a method of reacting cyclodextrin with an epoxy group or an isocyanate group at each terminal.

[0005]

However, in any of the above methods, since the silylating agent does not sufficiently react with the surface of the silica gel, etc., the loading amount is 1000 μmol / mol.
Silica gel is less than 1 g or less, and as a result, the fixed amount of cyclodextrin is 100 μmol / silica gel 1
It had the disadvantage of being very small, on the order of g or less.

Accordingly, an object of the present invention is to provide a method for immobilizing cyclodextrin on an inorganic substance, particularly silica gel, in which the amount of cyclodextrin immobilized on the inorganic substance is significantly increased.

[0007]

Means for Solving the Problems In view of the above problems, the present inventors have conducted intensive studies and as a result, the object of the present invention is to provide an allyloxycarboxylic acid derivative, an allyloxy alcohol derivative, and A compound represented by the following general formula [II] is reacted with cyclodextrin or a cyclodextrin derivative and then reacted with activated silica gel, or reacted with activated silica gel and then reacted with cyclodextrin or cyclodextrin derivative. A method for immobilizing cyclodextrin, characterized by reacting.

[0008]

Embedded image (Wherein, R ₁ represents a methoxy group, an ethoxy group, a chlorine atom, etc., R ₂ represents a methyl group, an ethyl group, etc., R ₃ represents a hydrogen atom, a methyl group, etc., and x ₃ represents 1 to ₃ . Represents an integer of 3, and y ₁ represents an integer of ₁ to 5.)

[0009]

Embedded image (Wherein R ₄ represents a methoxy group, an ethoxy group or the like;
₅ represents a methyl group, an ethyl group or the like. Further, x ₄ is 1
And y ₂ represents an integer of 1 to 5. )

Hereinafter, the present invention will be described more specifically.

The cyclodextrin (hereinafter abbreviated as CD) used in the present invention has a glucose unit of 6,
Any of 7 and 8 α, β and γ-cyclodextrins can be used.

The following are examples of CD derivatives used in the present invention. (1) α, having at least one hydroxyl group,
β, γ-CD derivatives. (2) at least one leaving group Α, β, γ-CD derivative having the formula: (3) α, β, γ-CD derivatives having at least one methacrylic acid type group (or acrylic acid type group).

The allyloxycarboxylic acid derivative and allyloxy alcohol derivative used in the present invention include:
Preferred are those represented by the following formulas.

[0014]

Embedded image (In the formula, x1 is an integer of ₁ to 5, and x2 is an integer of ₂ to 6.)

The "activated silica gel" used in the present invention refers to a silica gel in which a large number of hydroxyl groups are generated on the surface by refluxing silica gel in dilute nitric acid for one hour or more.

The method for immobilizing cyclodextrin according to the present invention comprises an allyloxycarboxylic acid derivative, an allyloxy alcohol derivative, an alkyldialkoxymethylsilane methacrylate or a 3-aminoalkyldialkoxymethylsilane (hereinafter referred to as "the present invention"). Derivatives)),
It is characterized by sequentially reacting CD or a CD derivative and activated silica gel. Such a reaction is carried out by reacting the derivative of the present invention with CD or CD derivative and then reacting with activated silica gel. Alternatively, it can be carried out by first reacting the derivative of the present invention with activated silica gel and then reacting it with CD or a CD derivative.

The following is a specific reaction example of the method for immobilizing CD of the present invention.

[0018]

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[0019]

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[0020]

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The above reaction can be specifically carried out as follows.

Reaction Example (1) Reaction [1] 3-Allyloxypropionic acid (x ₁ = 2) is reacted with a methanol solution of 1M-KOH in methanol to terminate the reaction at the neutralization point. Solidify.
The obtained white solid is recrystallized from isopropyl alcohol to obtain [A]. ([A] M; K, yield: about 85
%)

Reaction [2] 3-Allyloxypropionic acid (x ₁ = 2) is reacted in thionyl chloride at reflux for 6 hours. After completion of the reaction, thionyl chloride is distilled off under reduced pressure, and the residue is distilled under reduced pressure to obtain [B]. ([B] yield: about 80%)

Reaction [3] [A] is dissolved in dehydrated DMF (or DMSO) and heated to 60 ° C. under a nitrogen atmosphere. Then, a monotosyl (or monoiod) β-CD derivative (n = 7, R
₁ ; A DMF solution of CH ₃ CO-) is added dropwise. After completion of the dropwise addition, the mixture is stirred at 100 ° C. for 24 hours. After completion of the reaction, DMF is distilled off under reduced pressure, and the residue is extracted with methylene chloride / water. The methylene chloride phase is dried and concentrated, and the residue is purified by silica gel column chromatography to obtain [C]. ([C] yield:
About 55%)

Reaction [4] Monohydroxy β-CD derivative (n = 7, R ₁ ; CH ₃
-) Is dissolved in dehydrated THF, triethylamine is added, and the temperature is lowered to 0 to 5 ° C under a nitrogen stream. Thereafter, [B] is slowly added dropwise, and after completion of the addition, the mixture is stirred at 0 to 5 ° C. for 2 hours and at room temperature for 12 hours. After completion of the reaction, THF is distilled off under reduced pressure, and the residue is extracted with methylene chloride / water. The methylene chloride phase is dried and concentrated, and the residue is purified by silica gel column chromatography to obtain [C]. ([C] yield: about 75%)

Reaction [5] 3-allyloxypropionic acid (x ₁ = 2) and a monohydroxy β-CD derivative (n = 7, R ₁ = CH _3- ) are dissolved in dehydrated chloroform, and Add 4-pyrrolidinopyridine. Further, dicyclohexylcarbodiimide is added at room temperature, and the reaction is performed under reflux for 24 hours. After completion of the reaction, filtration was performed, and chloroform was concentrated under reduced pressure.
The residue is extracted with methylene chloride / water. The methylene chloride phase is dried and concentrated, and the residue is purified by silica gel column chromatography to obtain [C]. ([C] yield: about 40
%)

Reaction [6] [C] (x ₁ = 2, R ₁ ; CH ₃ CO—) and triethoxysilane (y = 3, R ₂ ; C ₂ H ₅ O—) are dissolved in THF, Stir under an argon atmosphere. A THF solution of hexachloroplatinic acid hexahydrate is slowly added dropwise to the system at room temperature, and after completion of the addition, the mixture is stirred at 80 ° C. for 24 hours. After completion of the reaction, the precipitate is removed by centrifugation, THF is distilled off under reduced pressure, and the residue is thoroughly dried. Since the obtained [D] is easily decomposed, it is stored under a nitrogen atmosphere. ([D] yield: about 80
%)

Reaction [7] The activated silica gel is dispersed in dehydrated toluene, and [D] (R ₂ ; C ₂ H ₅ O—, y = 3, x
₁ = 2, R ₁ ; CH ₃ CO—) and then 100 ° C.
And stir for 12 hours. After completion of the reaction, the reaction silica gel is separated by filtration, washed well with acetone, and dried at 80 to 90 ° C. under reduced pressure to obtain [E]. ([E] fixed amount: about 550 μmol / g silica gel)

Reaction Example (2) Reaction [8] 3-Allyloxypropionic acid (x ₁ = 2) is stirred in THF, and pyridine is further added. Then at room temperature in THF
Was added dropwise, and the mixture was stirred under reflux for 12 hours. After completion of the reaction, the mixture is filtered, THF pyridine is distilled off under reduced pressure, and the residue is recrystallized from benzene / chloroform to obtain [F]. ([F] yield: about 70%)

Reaction [9] [F] (x ₁ = 2, R ₃ ; (C ₆ H ₅ ) ₃ C—) and triethoxysilane (y = 3, R ₂ ; C ₂ H ₅ O—) Reaction According to the reaction [6]. ([G] yield: about 90%)

Reaction [10] [G] (x ₁ = 2, R ₃ ; (C ₆ H ₅ ) ₃ C—, y =
3, Reaction of R ₂ ＣC ₂ H ₅ O—) with activated silica gel According to the reaction [7]. ([H] supported amount: about 2200 μmol /
Silica 1g)

Reaction [11] [H] is dispersed in an aqueous acetone / HBr solution and
Stir for 30 minutes. After the completion of the reaction, the reaction silica gel was separated by filtration.
It is washed well with acetone and dried at 80 to 90 ° C. under reduced pressure to obtain [I]. ([H] → [I] conversion rate about 100%)

Reaction [12] [I] is dispersed in SOCl ₂ and stirred under reflux for 12 hours. After completion of the reaction, the reaction silica gel is separated by filtration, washed well with dehydrated acetone, and dried at 80 to 90 ° C. under reduced pressure [J].
Get. ([I] → [J] conversion rate about 100%)

Reaction [13] [J] was dispersed in dehydrated THF, triethylamine was added under a nitrogen atmosphere, and the monohydroxy β-CD derivative (n = 7, R ₁ = CH _3-) dissolved in THF at 0 to 5 ° C. ) Is dropped, and the mixture is allowed to react at 0 to 5 ° C for 2 hours and at room temperature for 24 hours after completion of the dropping. After completion of the reaction, the reaction silica gel is filtered off, and the residue is refluxed in methanol to crush the terminal reactive groups (about 12 hours). After completion, the reaction silica gel is filtered off, and 80-90 under reduced pressure.
[K] is obtained by drying at ° C. ([K] CD fixed amount: about 500 μmol / g of silica)

Reaction [14] [A] (x ₁ = 2, M; K) and triethoxysilane (y
_{= 3, R 2; C 2} H 5 O-) was dissolved in DMF, and stirred under an argon atmosphere. A DMF solution of hexachloroplatinic acid hexahydrate is slowly added dropwise to the system at room temperature, and after completion of the addition.
Stir at 80 ° C. for 24 hours. After completion of the reaction, the precipitate is removed by centrifugation, DMF is distilled off under reduced pressure, and the residue is thoroughly dried. Since the obtained [L] is easily decomposed, it is stored under a nitrogen atmosphere. ([L] yield: about 70%)

Reaction [15] [L] (x ₁ = 2, M; K, R ₂ ; C ₂ H ₅ O—, y =
Reaction of 3) with activated silica gel According to reaction [7]. ([M] supported amount: about 2000 μmol /
Silica 1g)

Reaction [16] [M] (x ₁ = 2, M; K, R ₂ ; C ₂ H ₅ O—) and S
Reaction with OCl _{2 According} to the reaction [12]. ([M] → [J] Conversion rate about 100
%)

Reaction [17] [M] (x ₁ = 2, M; K, R ₂ ; C ₂ H ₅ O-)
HF, and dispersed at room temperature under a nitrogen atmosphere at room temperature with a monotosyl (or monoiod) β-CD derivative (n = 7, R ₁ ;
A THF solution of CH ₃ CO—) is added dropwise. After dropping,
The reaction is performed for 48 hours under reflux. After completion of the reaction, the reaction silica gel is separated by filtration, washed well with hot water, and dried at 80 to 90 ° C. under reduced pressure to obtain [K]. ([K] CD fixed amount: about 300μ
Mol / silica gel 1g)

Reaction Example (3) Reaction [18] 3-Allyloxypropionic acid (x ₁ = 2) is reacted in thionyl chloride under reflux for 6 hours. After completion of the reaction, thionyl chloride is distilled off under reduced pressure, and the residue is distilled under reduced pressure to obtain [N]. ([N] yield: about 80%)

Reaction [19] [N] and trimethoxysilane (R ₁ ; CH ₃ O—, y =
3) is dissolved in THF and stirred under an argon atmosphere.
A THF solution of hexachloroplatinic acid hexahydrate is slowly added dropwise to the system at room temperature, and the mixture is stirred at 80 ° C. for 24 hours after completion of the addition.
After completion of the reaction, the precipitate was removed by centrifugation, and THF was added.
Is distilled off under reduced pressure to obtain a residue [O]. Since the compound is easily decomposed, it is stored under a nitrogen atmosphere. ([O] yield:
About 70%)

Reaction [20] Activated silica gel is dispersed in dehydrated toluene, and a toluene solution of [O] is added at room temperature under a nitrogen atmosphere.
React at 12 ° C for 12 hours. After completion of the reaction, the reaction silica gel is separated by filtration, washed well with toluene, and dried at 80 to 90 ° C. under reduced pressure to obtain [P]. ([P] fixed amount: about 1500 μmol / g silica gel)

Reaction [21] (in the case of [a]) [P] is dispersed in dehydrated pyridine, and the system is cooled to 0 to 5 ° C. under a nitrogen atmosphere. A pyridine solution of β-CD is slowly added dropwise to the system. After completion of the dropwise addition, the mixture is reacted at 0 to 5 ° C. for 2 hours and then at room temperature for 12 hours. After completion of the reaction, the reaction silica gel is separated by filtration, washed well with DMF, and refluxed in methanol (about 12 hours). After completion, the reaction silica gel is separated by filtration and dried at 80 to 90 ° C. under reduced pressure to obtain [Q]. (Amount of CD fixed to [P]: about 350 μmol / g of silica gel)

Reaction [21] (in the case of [b]) (R ₂ ; CH
₃ CO-, m = 7) In the reaction [21] [a], dehydrated pyridine is replaced with dehydrated THF.
/ Triethylamine and the β-CD pyridine solution
[Q] is obtained in the same manner except that a THF solution of -CD is used. (Amount of CD fixed to [P]: about 450 μmol / g of silica gel)

[0044]

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[0045]

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[0046]

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Reaction Example (4) Reaction [1] A THF solution of allyloxyethanol (x ₂ = 2) is added dropwise to an aqueous sodium hydroxide solution at 0 to 5 ° C., followed by stirring at 0 ° C. or lower for 1 hour. Next, a THF solution of p-toluenesulfonic acid chloride is added dropwise, and the mixture is stirred at 0 ° C. or lower for 3 hours even after completion of the addition. After completion of the reaction, the reaction solution is separated, the aqueous phase is extracted with methylene chloride, combined with the THF phase, dried and concentrated under reduced pressure, and the residue is purified by silica gel column chromatography to obtain [A]. ([A] yield: about 85%)

Reaction [2] NaH is dispersed in THF under a nitrogen stream, and a THF solution of a monohydroxy β-CD derivative (n = 7, R ₂ ; CH ₃ —) is added dropwise at room temperature. After completion of the dropwise addition, the mixture was stirred for 2 hours, and then a THF solution of [A] (x ₂ = 2) was slowly added dropwise.
After the completion of the dropwise addition, the reaction is carried out under reflux for 24 hours. After completion of the reaction, the reaction mixture is filtered, the filtrate is distilled off under reduced pressure, the residue is extracted from methylene chloride / water, and the methylene chloride phase is dried and concentrated under reduced pressure. The residue is purified by silica gel column chromatography to obtain [B]. ([B] yield: about 55%)

Reaction [3] [B] (x ₂ = 2, n = 7, R ₂ ; CH _3- ) and dimethoxymethylsilane (y = 2, R ₃ ; CH ₃ O--) are reacted with TH.
Dissolve in F and stir under an argon atmosphere. A THF solution of hexachloroplatinic acid hexahydrate is slowly added dropwise to the system at room temperature, and the mixture is stirred at 80 ° C. for 24 hours after completion of the addition. After completion of the reaction, the precipitate is removed by centrifugation, THF is distilled off under reduced pressure, and the residue is thoroughly dried. Since the obtained [C] is easily decomposed, it is stored under a nitrogen atmosphere. ([C] yield: about 85%)

Reaction [4] The activated silica gel is dispersed in dehydrated toluene, and [C] (R ₃ ; CH ₃ O—, y = 2, x ₂ at room temperature in a nitrogen atmosphere.
= 2, n = 7, R ₂ ; CH ₃ —), and then 100
Stir at 12 ° C. for 12 hours. After completion of the reaction, the reaction silica gel is separated by filtration, washed well with acetone, and dried at 80 to 90 ° C. under reduced pressure to obtain [D]. ([D] fixed amount: about 600 µmol / g silica gel)

Reaction [5] Allyloxyethanol (x ₂ = 2) is dissolved in dehydrated THF, and triethylamine is further added. The system was cooled to 0 to 5 ° C. under a nitrogen atmosphere, and a THF solution of a monoacidified β-CD derivative (n = 7, R ₁ ; CH ₃ CO—) was slowly added dropwise thereto. For 2 hours and then at room temperature for 6 hours. After completion of the reaction, THF and triethylamine are distilled off under reduced pressure, the residue is extracted from methylene chloride / water, the methylene chloride phase is dried and concentrated under reduced pressure, and the residue is purified by silica gel column chromatography to obtain [E]. ([E] yield: about 75%)

Reaction [6] Reaction of [E] (x ₂ = 2, n = 7, R ₁ ; CH ₃ CO—) with dimethoxymethylsilane (R ₃ ; CH ₃ O—, y = 2) 3]. ([F] yield: about 80%)

Reaction [7] [F] (x ₂ = 2, n = 7, R ₁ ; CH ₃ CO—, R
₃ ; Reaction between CH ₃ O—, y = 2) and activated silica gel According to the reaction [4]. ([G] fixed amount: about 550 μmol /
Silica 1g)

Reaction Example (5) Reaction [8] Allyloxyethanol (x ₂ = 2) in dehydrated pyridine
Is dissolved and trityl chloride is added at room temperature. Then 60
Incubate for 3 hours at ° C. After completion of the reaction, pyridine is distilled off under reduced pressure, the residue is extracted from ether / water, and the ether phase is dried and concentrated under reduced pressure. The residue is purified by silica gel column chromatography to obtain [H]. ([H] yield: about
90%)

Reaction [9] Reaction of [H] (x ₂ = 2, R ₄ ; (C ₆ H ₅ ) ₃ C—) with methyltrichlorosilane (R ₃ ; Cl, y = 3) Reaction [3] According to. ([I] yield: about 75%)

Reaction [10] [I] (x ₂ = 2, R ₄ ; (C ₆ H ₅ ) ₃ C—, R ₃ ;
Reaction of activated silica gel with Cl, y = 3) According to the reaction [4]. ([J] supported amount: about 2000 μmol /
Silica gel 1g)

Reaction [11] [J] was dispersed in an acetone / HBr aqueous solution, and
Stir for 30 minutes. After the completion of the reaction, the reaction silica gel was separated by filtration.
It is washed well with acetone, and dried at 80 to 90 ° C. under reduced pressure to obtain [K]. ([J] → [K] conversion rate about 100%)

Reaction [12] [K] (x ₂ = 2) is dispersed in dehydrated THF, triethylamine is added under a stream of nitrogen, and the system is cooled to 0 to 5 ° C. A monoacidified β-CD derivative (n = 7, R
₅ ; A THF solution of CH _3- ) is slowly added dropwise. After completion of the dropwise addition, the mixture is reacted at 0 to 5 ° C for 2 hours and at room temperature for 24 hours. After completion of the reaction, the reaction silica gel was separated by filtration, and the residue was thoroughly washed with methanol and dried at 80 to 90 ° C. under reduced pressure to obtain [L].
Get. ([L] CD fixed amount: about 600 μmol / g silica gel)

Reaction Example (6) Reaction [13] A THF solution of allyloxyethanol (x ₂ = 2) is added dropwise to an aqueous sodium hydroxide solution at 0 to 5 ° C., and the mixture is stirred at 0 ° C. or lower for 1 hour. Then, a THF solution of p-toluenesulfonic acid chloride was added dropwise, and after the addition was completed,
Stir below 3 ° C for 3 hours. After completion of the reaction, the reaction solution is separated, the aqueous phase is extracted with methylene chloride, combined with the THF phase, dried and concentrated under reduced pressure, and the residue is purified by silica gel column chromatography to obtain [M]. ([M] yield: about 85%)

Reaction [14] [M] and sodium iodide are stirred in acetone for 12 hours under reflux. After completion of the reaction, the mixture is filtered, and the acetone is concentrated under reduced pressure. The residue is purified by silica gel column chromatography to obtain [N]. ([N] yield: about 90%)

Reaction [15] [N] and triethoxysilane (R ₁ ; C ₂ H ₅ O—, y
= 3) is dissolved in THF and stirred under an argon atmosphere. At room temperature, the TH of hexachloroplatinic acid hexahydrate was added to the system.
The solution F is slowly added dropwise, and after completion of the addition, the mixture is stirred at 80 ° C. for 24 hours. After the reaction was completed, the precipitate was removed by centrifugation, and TH
F is concentrated under reduced pressure to obtain a residue [O]. Since the compound is easily decomposed, it is stored under a nitrogen atmosphere. ([O] yield:
About 80%)

Reaction [16] Activated silica gel is dispersed in dehydrated toluene, and a toluene solution of [O] is added at room temperature under a nitrogen atmosphere.
React at 12 ° C for 12 hours. After completion of the reaction, the reaction silica gel is separated by filtration, washed well with toluene, and dried at 80 to 90 ° C. under reduced pressure to obtain [P]. ([P] fixed amount: about 1800 μmol / g silica gel)

Reaction [17] (in the case of [a]) [P] is dispersed in dehydrated DMF, and a DMF solution of Na alcoholate of β-CD (m = 7) separately filtered at room temperature under a nitrogen atmosphere. Is dropped. After the dropwise addition, heat slowly and stir at 120 ° C for 24 hours. After completion of the reaction, the reaction silica gel was separated by filtration, washed well with DMF and water, and reduced under reduced pressure to 80 to 90.
[Q] is obtained by drying at ° C. (Amount of CD fixed to [P]: about 250 μmol / g of silica gel) In the case of γ-CD (m = 8), 12 ° C.
The reaction time is 12 hours at 80 ° C and 24 hours at 120 ° C.

Reaction [17] (in the case of [b]) (R ₂ ; CH
₃ −, m = 7) In the reaction [17] [a], DMF was THF,
[Q] is obtained in the same manner except that the reflux temperature is 120 ° C.
(Amount of CD derivative immobilized on [P]: about 400 μmol / g of silica gel)

Reaction [18] [R] is obtained in the same manner as in reaction [15] except that [N] is changed to [M].

Reaction [19] [S] is obtained in the same manner as in reaction [16] except that [O] is changed to [R].

Reaction [20] [Q] is obtained in the same manner as in reactions [17] [a] and [17] [b] except that [P] is changed to [S].

[0068]

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Reaction Example (7) Activated silica gel and propyldiethoxymethylsilane methacrylate are refluxed for 6 hours in dehydrated toluene in the presence of a small amount of hydroquinone. After the reaction is completed, the reaction silica gel is left to cool, and the reaction silica gel is filtered, washed with toluene and methanol, and heated to 80 to 90 ° C.
Dry under reduced pressure. Next, the reaction silica gel, a separately synthesized methacrylic ester type CD derivative, and a toluene solution of azobisisobutyronitrile as an initiator are put into a polymerization tube, and the tube is sealed after repeated freezing and deaeration. After sealing, the polymerization tube is shaken at 70-80 ° C for 24 hours. After the completion of the reaction, the mixture is left to cool and filtered. The residue is thoroughly washed with toluene and methanol, and dried under reduced pressure at 80 to 90 ° C. (CD fixed amount: about 500μmol / silica gel 1g)

[0070]

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Reaction Example (8) Activated silica gel is refluxed with 3-aminopropyldiethoxymethylsilane in dehydrated toluene for 3 hours. After completion of the reaction, the reaction mixture is left to cool, the reaction silica gel is filtered off, washed well with toluene and methanol, and dried. The resulting reaction silica gel is reacted with methacrylic anhydride in dehydrated methylene chloride at 0-5 ° C. for 1 hour and then at room temperature for 24 hours. After the reaction,
The reaction silica gel is separated by filtration and washed well with methylene chloride and methanol. Thereafter, drying is performed under reduced pressure at 80 to 90 ° C. Next, a DMF solution of the obtained reaction silica gel, CD derivative (methacrylic ester type), and azobisisobutyronitrile is put in a polymerization tube, and the tube is sealed after repeated freezing and deaeration. After sealing, the polymerization tube is shaken at 70-80 ° C for 24 hours.
After the completion of the reaction, the mixture is left to cool and filtered. The residue is thoroughly washed with DMF and methanol, and dried under reduced pressure at 80 to 90 ° C. (CD fixed amount: about 400 μmol / silica gel 1 g)

Using the stationary phase synthesized by the method for immobilizing CD of the present invention, the following 12 dansyl amino acids were optically resolved under the following conditions.

[0073]

Embedded image HPLC model used: Model-6000A manufactured by Waters Column content used Length: 150 mm, inner diameter: 4.6 mm (filled with a CD stationary phase in this column) HPLC measurement conditions Eluent: triethylamine and acetic acid buffer solution / methanol = 50/50 pH: 5.0 Flow rate:
1.0 ml / min The degree of optical resolution (that is, the degree of separation between R-form and L-form) is shown by percentage.

That is, the degree of separation was numerically expressed for two optical isomer peaks appearing on the chromatogram. That is, the height from the peak to the baseline is h
Assuming that the height from the peak to the bottom of the valley between the two peaks is h ′, the degree of separation R ′ is expressed by the following equation. R ′ (%) = h ′ / h × 100 Accordingly, the degree of separation is 0% when no separation is made, and 100% when two peaks are separated to the baseline. Table 1 shows the results.

[0075]

[Table 1] However, the stationary phase A was synthesized according to the method described in US Pat. No. 4,539,399, and the stationary phase B was synthesized according to the reaction example (6) of the present invention.

From the above results, it is clear that the newly synthesized stationary phase B is very excellent in optical resolution.

[0077]

According to the present invention, by immobilizing cyclodextrin on an inorganic solid, the inclusion effect can be utilized to provide an excellent insoluble separation / adsorbent.

In particular, by increasing the amount of the spacer carried on silica gel, the amount of cyclodextrin immobilized in the subsequent reaction can be increased, thereby greatly increasing the function of cyclodextrin.

Claims (4)

(57) [Claims] 1. An allyloxycarboxylic acid derivative is reacted with cyclodextrin or a cyclodextrin derivative and then with activated silica gel, or reacted with activated silica gel and then with cyclodextrin or a cyclodextrin derivative. A method for immobilizing cyclodextrin, comprising: 2. The reaction of an allyloxy alcohol derivative with cyclodextrin or a cyclodextrin derivative and then with activated silica gel, or with the activated silica gel and then with a cyclodextrin or cyclodextrin derivative. A method for immobilizing cyclodextrin, which is characterized in that: 3. The compound represented by the following general formula [I] is reacted with cyclodextrin or a cyclodextrin derivative and then with activated silica gel, or is reacted with activated silica gel and then cyclodextrin or cyclodextrin. A method for immobilizing cyclodextrin, which comprises reacting with a derivative. Embedded image (Wherein, R ₁ represents a methoxy group, an ethoxy group, a chlorine atom, etc., R ₂ represents a methyl group, an ethyl group, etc., R ₃ represents a hydrogen atom, a methyl group, etc., and x ₃ represents 1 to ₃ . Represents an integer of 3, and y ₁ represents an integer of ₁ to 5.) 4. A compound represented by the following general formula [II] is reacted with cyclodextrin or a cyclodextrin derivative and then with activated silica gel, or is reacted with activated silica gel and then cyclodextrin or cyclodextrin. A method for immobilizing cyclodextrin, which comprises reacting with a derivative. Embedded image (Wherein R ₄ represents a methoxy group, an ethoxy group or the like;
₅ represents a methyl group, an ethyl group or the like. Further, x ₄ is 1
And y ₂ represents an integer of 1 to 5. )