JPH0440660B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a chromatographic packing material grafted with a novel optically active organosilane, and an -NH- group, -OCNH- group,
There is a method for separating and analyzing an enantiomeric mixture of a compound having an -OCO- group or an -OH group by liquid chromatography. As a filler grafted with an optically active compound for directly separating and analyzing an enantiomeric mixture of a compound having an asymmetric carbon by liquid chromatography, for example, Davankov et al. grafted optically active proline. A method using a ligand exchange using a filler, a method using a charge transfer complex using a filler grafted with an optically active compound lacking π electrons by Gil-Av, etc., a method using a charge transfer complex using a filler grafted with an optically active N-acylated amino acid by Hara et al. Separation of N-acylated amino acid esters and N-acylated dipeptide esters using packing materials or Pirkle
3,5-dinitrobenzoylated amino acids using optically active 1-(9-anthryl)trifluoroethanol-grafted fillers by et al.
Separation of amines, oxyacids, sulfoxides, etc. and separation of aromatic alcohols using fillers grafted with 3,5-dinitrobenzoylated optically active phenylglycine have been reported. However, these methods are limited to a narrow range of compounds that can be separated, the degree of separation is small, and it is difficult to produce grafted fillers, making it difficult to produce fillers with reproducible performance. It is difficult to obtain a filler, and it is difficult to say that any of them are practical fillers. Under such circumstances, the present inventors have continued to conduct intensive studies with the aim of developing a grafted filler that is applicable to a wide range of compounds that can be analyzed, is relatively easy to manufacture, and is chemically stable and practical. As a result, optically active pheniglycine acylated with an optically active chrysanthemum acid or α-(4-chlorophenyl)isovaleric acid residue containing an asymmetric carbon was grafted onto an inorganic carrier having a hydroxyl group on its surface. -NH- group bonded to the asymmetric carbon, -
It not only shows excellent effects in separating enantiomeric mixtures of compounds having CONH-, -OCO-, or -OH groups, but also can be easily produced by standard chemical reactions and is chemically stable. We have discovered that these are extremely useful fillers, leading to the present invention. That is, the present invention combines optically active phenylglycine acylated with optically active chrysanthemum acid or α-(4-chlorophenyl)isovaleric acid containing an asymmetric carbon onto an inorganic carrier having a hydroxyl group on its surface. A chromatographic packing material grafted with an optically active organosilane formed by bonding an aminoalkylsilane, and -NH- bonded to an asymmetric carbon using the same as a stationary phase of liquid chromatography.
A method is provided for separating and analyzing enantiomeric mixtures of compounds having -CONH, -OCO- or -OH groups. The present invention will be described in further detail. As the organosilane grafted in the present invention, for example, the general formula [] [In the formula, R ₁ , R ₂ and R ₃ are selected from an alkyl group, an alkoxyl group, a hydroxyl group, or a halogen atom, and at least one is an alkoxyl group or a halogen atom. n is an integer from 2 to 4. A is optically active 3-(2-methyl-1
-propenyl)-2,2-dimethyl-1-cyclopropyl group or optically active 1-(4-chlorophenyl)-isobutyl group. * represents an asymmetric carbon] Optically active N-acylated phenylglycine derivatives can be mentioned. Further, as the aminoalkylsilane component, ω-aminoalkylalkoxysilane or ω-aminoalkylhalogenosilane is preferable, and examples thereof include γ-aminopropyltriethoxysilane and γ-aminopropyltrichlorosilane. In the present invention, the inorganic carrier having hydroxyl groups on its surface is preferably a silica-containing carrier such as silica gel, and the carrier may have any shape such as spherical or crushed. In order to obtain this, it is preferable to use fine particles with as uniform a particle size as possible. Various grafting methods can be employed to prepare the novel chromatographic packing material of the present invention, including the following methods. An inorganic carrier having a hydroxyl group on its surface is reacted with an aminoalkylsilane to introduce an aminoalkylsilyl residue onto the surface of the inorganic carrier,
A method of reacting this with optically active phenylglycine acylated with an optically active chrysanthemum acid or α-(4-chlorophenyl)isovaleric acid residue to cause dehydration condensation. More specifically, an inorganic carrier having a hydroxyl group on its surface has the general formula [] [In the formula, R ₁ , R ₂ , R ₃ , and n have the same meanings as above. ] The aminoalkylsilane represented by the formula [] is reacted by a known method to introduce an aminoalkylsilyl residue onto the surface of the inorganic carrier, and then the general formula [] [wherein A and * have the same meanings as above. ] An optically active N-acylated phenylglycine having an asymmetric carbon represented by, for example, N-chrysanthemoylphenylglycine, N-{α-(4-chlorophenyl)isovaleroyl}phenylglycine, etc. is reacted, The desired filling can be obtained by dehydration condensation. In addition, optically active N- having an asymmetric carbon shown in the above general formula []
Acylated phenylglycine can be synthesized by commonly used methods, such as primary chrysanthemum acid or α
-(4-chlorophenyl)isovaleric acid is used as its acid chloride, and it is obtained by reacting the acid chloride with pheniglycine in the presence of a dehydrochlorination agent. A method in which an organosilane obtained by reacting optically active N-acylated phenylglycine with an aminoalkylsilane is grafted onto an inorganic carrier having a hydroxyl group on its surface. More specifically, N-acylated phenylglycine represented by the general formula [] is combined with the general formula []
The desired filler can be obtained by grafting an organosilane represented by the general formula [] obtained by reacting an aminoalkylsilane represented by the following onto an inorganic carrier such as silica gel. The packing material having an optically active N-acylated phenylglycine residue obtained according to the present invention is packed into a chromatography column according to a conventional method and used as a stationary phase in liquid chromatography. In liquid chromatography using this stationary phase, by selecting appropriate elution conditions, especially the commonly used normal phase partition conditions, -
NH- group, -CONH- group, -OCO- group or -OH
Separation and analysis of a mixture of enantiomers of a compound having a group can be performed with good resolution and in a short time. Example 1 Silica gel (particle size 10 μm, pore size 60 Å, surface area 500
m ² /g) 10g was dried under reduced pressure at 130°C for 4 hours, and then 20g of 3-aminopropyltriethoxysilane was added.
Add to 200ml of dehydrated toluene solution and heat at 60℃.
Stir for 6 hours. The reaction mixture was filtered, and the residue was washed with 100 ml of acetone and dried to obtain 3-aminopropyl silylated silica gel (APS). The elemental analysis values of this product were N: 1.20% and C: 3.40%, which corresponds to about 0.90 mmol of 3-aminopropyl group grafted to 1 g of silica gel. Separately, dissolve 10 g of D-phenylglycine in 33 ml of 2N sodium hydroxide solution, add 20 ml of ethyl ether, and while stirring vigorously under ice-cooling, dissolve 15 g of (+)-trans-chrysanthemum acid chloride and 40 ml of 2N sodium hydroxide solution. Add in 7 portions, approximately every 15 minutes. After stirring vigorously for another 2 hours at room temperature, the reaction solution was washed twice with 50 ml of ethyl ether, acidified with 6N hydrochloric acid, and the resulting oil was washed with 100 ml of ethyl acetate.
Extract 3 times. Wash the extract twice with 100ml of water,
After dehydration over anhydrous sodium sulfate and concentration under reduced pressure, recrystallization from a mixture of ethyl acetate and n-hexane yielded N-(+)-trans-chrysanthemoyl-D.
-15 g of phenylglycine was obtained as white crystals. Melting point: 76-80℃ Optical rotation: [α] ²⁰ _D = -38.8゜ (c = 2%, chloroform) Elemental analysis values Carbon (%) Hydrogen (%) Nitrogen (%) Calculated value 71.74 7.70 4.65 Actual value 71.58 7.65 4.58 Next, 12 g of this compound and 4.6 g of N-hydroxysuccinimide are dissolved in 100 ml of dehydrated tetrahydrofuran, and while cooling on ice and stirring vigorously, 8.3 g of dicyclohexylcarbodiimide is dissolved in 20 ml of dehydrated tetrahydrofuran and added dropwise over 30 minutes. After stirring for 2 hours under ice-cooling and further stirring for 3 hours at room temperature, the precipitate was removed by filtration, and the solvent was distilled off under reduced pressure. The residue was dissolved in 150 ml of ethyl acetate, washed twice with 50 ml of water, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and then recrystallized from a mixture of ethyl acetate and n-hexane to obtain the formula 12 g of the compound represented by was obtained as white crystals. Melting point: 80-85℃ Optical rotation: [α] ²⁰ _D = 122.9゜ (c = 2%, chloroform) Elemental analysis values Carbon (%) Hydrogen (%) Nitrogen (%) Calculated value 66.32 6.58 7.03 Actual value 66.53 6.31 6.99 Next, take 6 g of this compound, suspend 3 g of the aminopropyl silylated silica gel (APS) in 30 ml of dehydrated tetrahydrofuran, add to the sufficiently degassed liquid under reduced pressure, and heat at room temperature for 5 hours, then at 50°C.
Stir gently for 5 hours. After cooling to room temperature, the desired filler was washed three times with 30 ml of tetrahydrofuran, then twice with 30 ml of methanol, and then twice with 30 ml of ethiether, dried, and grafted with N-(+)-trans-chrysanthemoyl-phenylglycine. I got it. The elemental analysis value of this item is N: 1.74
%, C: 13.3%, which is N-(+)-trans-chrysanthemoyl-D per 1 g of silica gel.
- indicates that approximately 0.53 mmol of phenylglycine was grafted. The thus obtained packing material was slurried packed into a stainless steel column with an inner diameter of 4 mm and a length of 30 cm, and (±)-1-(4-chlorophenyl)-4,4-dimethyl-2-( 1, 2, 4
-triazol-1-yl)-1-penten-3
-ol was analyzed and the chromatogram shown in Figure 1 was obtained. Temperature: room temperature Mobile phase: hexane/1,2-dichloroethane/
Ethanol (350:40:4) Flow rate: 1.0ml/min Detector: Ultraviolet absorption meter (wavelength 254nm) In Figure 1, peak number (1)(-)-1-(4-chlorophenyl)-4,4- Dimethyl-2-(1,2,
4-triazol-1-yl)-1-pentene-
3-ol, (2) is (+)-1-(4-chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-yl)-1-penten-3-ol. each peak. The time required for peak (1) to elute is approximately 14 minutes, the separation factor is 1.22, and the time required for peak (1) and (2) to elute is approximately 14 minutes.
The area ratio of the peaks was 50:50. Example 2 (+)-α-(4-chlorophenyl)isovaleric acid
17.0g in n-hexane/toluene mixture (6:1)
Dissolve in 70ml, add 9mg of dimethylforma and add 40ml
Warm to ℃. To this, 11.5 g of thionyl chloride was added dropwise over 1 hour, and after stirring at 40°C for 5 hours, the solvent was distilled off under reduced pressure at room temperature, resulting in approximately 17.5 g of (+)-α-(4-clofeny)isovaleric acid chloride. get. Dissolve 9.5 g of D-phenylglycine in 32 ml of 2N sodium hydroxide solution, add 20 ml of ethyl ether, and stir vigorously under ice cooling to dissolve the above (+)
-Approximately 17.5 g of α-(4-chlorophenyl)isovaleric acid chloride and 35 ml of 2N sodium hydroxide solution in 10
After addition in 7 portions at ~15 minute intervals and stirring vigorously at room temperature for another 2 hours, the reaction mixture was washed twice with 50 ml of ethyl ether, acidified with 6N hydrochloric acid, and the resulting oil was diluted with 100 ml of ethyl acetate for 30 minutes. Extract times. The extract was washed twice with 100 ml of water, dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure. After adding 200 ml of n-hexane to the residue, stirring and washing, the lower layer was taken and the solvent was distilled off under reduced pressure. 21 g of the compound represented by was obtained. Melting point: 68-70℃ Optical rotation: [α] ²⁰ _D = -105.2゜ (c = 2.2%, tetrahydrofuran) Elemental analysis values Carbon (%) Hydrogen (%) Nitrogen (%) Chlorine (%) Calculated value 65.99 5.83 4.05 10.25 Actual value 65.49 5.99 3.79 10.41 Next, take 4 g of this compound, dissolve it in 40 ml of dehydrated tetrahydrofuran, and add 1.5 g of this compound while stirring under ice cooling.
-ethoxycarbonyl-2-ethoxy-1,2-
After adding 3.2 g of dihydroquinoline and stirring for 1 hour under ice cooling, 2.5 g of 3-aminopropylsilylated silica gel (APS) obtained in Example 1 was added.
After thoroughly deaerating under reduced pressure, the mixture was gently stirred at room temperature for one day and night. The reaction mixture was washed 4 times with 30 ml of tetrahydrofuran, then twice with 30 ml of methanol, and then twice with 30 ml of ethyl ether, dried, and N-{(+)-
α-(4-chlorophenyl)isovaleroyl}-D
- A desired filler grafted with phenylglycine was obtained. The elemental analysis value of this item is N: 1.26%,
C: 8.72%, which is N- per 1g of silica gel.
This shows that about 0.34 mmol of {(+)-α-(4-chlorophenyl)isovaleroyl}-D-phenylglycine was grafted. The thus obtained packing material was slurried packed into a stainless steel column with an inner diameter of 4 mm and a length of 25 cm.
N-3,5-dinitrobenzoyl-
(R,S)-1-cyclohexylethylamine was analyzed and the chromatogram shown in Figure 2 was obtained. Temperature: Room temperature Mobile phase: n-hexane/1,2-dichloroethane/ethanol (100:20:1) Flow rate: 1.0ml/min Detector: Ultraviolet absorption meter (wavelength: 254nm) Peak number (1) in Figure 2 ) is N-3,5-dinitrobenzoyl-(S)-1-cyclohexylethylamine, (2) is N-3,5-dinitrobenzoyl-(R)
-1-Cyclohexylethylamine peaks. The time required for peak (1) to elute was approximately 10 minutes, the fraction coefficient was 1.20, and the area ratio of peaks (1) and (2) was 50:50. Comparative Example N-(+)-trans-chrysanthemoyl-D of the present invention obtained in Examples 1 and 2, respectively
-Silica gel grafted with phenylglycine (hereinafter abbreviated as CHR-PHG-Si), N-{(+)-α
-(4-chlorophenyl)isovaleroyl}-D-
Silica gel grafted with phenylglycine (hereinafter abbreviated as CPI-PHG-Si) and JP-A-56-1350
N-formyl- produced according to Example 1 of the publication
Silica gel grafted with L-valine (hereinafter referred to as
FVA) was packed into a stainless steel column with an inner diameter of 4 mm and a length of 25 cm as a slurry, and the enantiomers of the following compounds were analyzed under the following conditions and the separation performance of each was compared. The inventive filler is
N-3,5-dinitrobenzoylated amino acid ester, N-acylated amine compared to FVA,
Excellent performance was demonstrated in the separation of enantiomers of carboxylic acid anilides. Temperature: Room temperature Flow rate: 1.0ml/min Detector: Ultraviolet absorption meter (wavelength: 254nm) [Table]

[Brief explanation of drawings]

Figure-1 and Figure-2 are Examples 1 and 2, respectively.
This is a chromatogram obtained in , where the vertical axis represents intensity and the horizontal axis represents retention time.

Claims (1)

[Claims] 1. General formula [] [In the formula, R ₁ , R ₂ and R ₃ are selected from an alkyl group, an alkoxyl group, a hydroxyl group, or a halogen atom, and at least one is an alkoxyl group or a halogen atom. n is an integer from 2 to 4. A is optically active 3-(2-methyl-1
-propenyl)-2,2-dimethyl-1-cyclopropyl group or optically active 1-(4-chlorophenyl)-isobutyl group. A chromatographic packing material in which an optically active organosilane represented by *represents an asymmetric carbon is grafted onto an inorganic carrier having hydroxyl groups on its surface. 2. The chromatographic packing material according to claim 1, wherein the inorganic carrier having hydroxyl groups on its surface is silica gel. 3 In the above general formula [], R ₁ , R ₂ and R ₃
The chromatographic packing material according to claim 1 or 2, wherein is an ethoxy group and n is 3. 4 In the above general formula [], R ₁ , R ₂ and R ₃
The chromatographic packing material according to claim 1 or 2, wherein is Cl and n is 3. 5 General formula [] [In the formula, R ₁ , R ₂ and R ₃ are selected from an alkyl group, an alkoxyl group, a hydroxyl group, or a halogen atom, and at least one is an alkoxyl group or a halogen atom. n is an integer from 2 to 4. A is optically active 3-(2-methyl-1
-propenyl)-2,2-dimethyl-1-cyclopropyl group or optically active 1-(4-chlorophenyl)-isobutyl group. An optically active organosilane represented by *represents an asymmetric carbon is grafted onto an inorganic support having a hydroxyl group on its surface. Using a chromatographic packing material, an -NH- group is bonded to an asymmetric carbon. , −CONH−
A liquid chromatography analysis method characterized by separating and analyzing a mixture of enantiomers of a compound having a -OCO- group or an -OH group. 6. The analytical method according to claim 5, which uses a chromatographic packing material in which the inorganic carrier having hydroxyl groups on its surface is silica gel. 7 In the above general formula [], R ₁ , R ₂ and R ₃
The analytical method according to claim 5 or 6, which uses a chromatographic packing material in which is an ethoxy group and n is 3. 8 In the above general formula [], R ₁ , R ₂ and R ₃
The analytical method according to claim 5 or 6, which uses a chromatographic packing material in which is a Cl group and n is 3.