JPS5950358A - Chromatograph filler grafted with optical active carboxylic acid and separation of enantiomer mixture using chromatograph filler - Google Patents

Abstract

PURPOSE:To obtain a chromatograph filler which is relatively easy to manufacture, is chemical stable and is practicable by grafting an optically active carboxylic acid having a specific formula. CONSTITUTION:A chromatograph filler wherein an optical active carboxylic acid deriv. expressed by the general formula I is grafted to an inorg. carrier having a hydroxyl group on its surface is obtd. by the method of bringing the aminoalkylsilane expressed by the general formula II into reaction by a known method with an inorg. carrier having a hydroxyl group on its surface to introduce a residual aminoalkylsiryl on the surface of the inorg. carrier then subjecting the same to dehydration condensation with an optically active carboxylic acid by using a condensation reagent such as N, N'-dicyclohexylcarbo di-imide, 1-ethoxycarbonyl-2-ethoxy-1,2-dehydroquinoline or the like.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention is directed to a novel chromatographic filler grafted with an optically active carboxylic acid, and one CONH- group, -0CO- group, -OH group, -0C- group bonded to an asymmetric carbon using the same.
The present invention relates to a method for separating and analyzing an enantiomeric mixture of a compound having an ONH- group or an -NCQNH- group by liquid chromatography.

To date, only a small number of packing materials grafted with optically active compounds have been reported for the direct separation and analysis of enantiomeric mixtures of compounds having asymmetric carbon atoms by liquid chromatography. For example, a method using a ligand exchange method using a filler grafted with optically active proline by Davankov et al., a method using a charge transfer complex using a filler grafted with an optically active compound lacking π electrons such as G11-Ay, an optical method using a layer, etc. Separation of active N-acylated amino acid esters and N-acylated dipeptide esters, or using a packing material grafted with optically active 1-(9-anthryl)trifluoro-dethanol by Pirk1e etc.3,5 Examples include the separation of dinitrobenzoylated amino acids, amines, oxyacids, sulfoxides, etc., and the separation of aromatic alcohols using fillers grafted with 8,5-dinitrobenzoylated optically active phenylglycine.

However, these methods are limited to a narrow range of compounds that can be separated, the degree of separation is small, and furthermore, it is difficult to produce grafted fillers, making it difficult to obtain fillers with reproducible performance. Therefore, it is difficult to say that any of them are practical fillers.

Under such circumstances, the present inventors have conducted intensive studies with the aim of developing a practical grafted filler that is applicable to a wide range of compounds that can be analyzed, is relatively easy to manufacture, and is chemically stable. As a result of continuing, the general formula [IJ R2-? i (CH2)1-NH-CO-R4(
Ill 3 [wherein R,, n2 and R3 are the same or different,
represents an alkyl group, an alkoxyl group, a hydroxyl group or a halogen atom, and at least one of Rt, R2a and R3 is an alkoxyl group or a halogen atom. n is an integer from 2 to 4, R4-(
:0 represents various carboxylic acid residues, and R4 has one or more asymmetric carbon atoms. ] A chromatographic filler grafted to an inorganic support having a carboxylic acid derivative carboxylic acid derivative carboxylic group on its surface or a -CON H- group bonded to an asymmetric carbon, Q
CO-A! ,, 1 of the enantiomeric mixture of compounds having -OH group, -0CONiI- group or -NCONH- group
The present inventors have discovered that it is an extremely useful filler that not only exhibits excellent effects on 13 compounds, but also can be easily produced by the chemical reaction described in 7i~ and is chemically stable, leading to the present invention. It is something.

Further details regarding the invention are described below.

In the carboxylic acid derivative represented by the above general formula [1], C as the carboxylic acid residue may be an α-arylalkylcarboxylic acid residue or a cyclopropanecarboxylic acid residue, and α-arylalkylcarboxylic acid component as,
Preferred are 1-lower alkylcarboxylic acids substituted with an aromatic group at the α-position, such as 2-(4-chlorophenyl)isovaleric acid and 2-phenylpropionic acid. C1 may include, for example, primary acids.

In addition, as the aminoalkylsilane component, ω-aminoalkylalkoxysilane or ω-aminoalkylhalogensilane is preferable, such as ω-aminopropyltriethoxysilane, ω-aminopropyltrichlorosilane, etc. 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 shape of the carrier may be either spherical or crushed. In order to achieve this, it is preferable to use fine particles that are as uniform in particle size as possible.

In preparing the novel romatograph filler of the present invention, various grafting methods can be employed, including, for example, the following methods.

(C) A method in which an inorganic carrier having a hydroxyl group on its surface is reacted with an aminoalkylsilane, an aminoalkylsilyl residue is introduced onto the surface of the inorganic carrier, and an optically active carboxylic acid is reacted with this.

J, more specifically, an inorganic carrier having a hydroxyl group on its surface has the general formula (Kawa I R2-5i -(CH+)n-NH2 ] The aminoalkylsilane represented by is reacted by a known method, an aminoalkylsilyl residue is introduced onto the surface of an inorganic carrier, and then an optically active carboxylic acid is added to the N,N-
A method of dehydration condensation using a condensation reagent such as dicyclohexyl carposi f-mide and l-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline, and a condensation method using an optically active carboxylic acid as an active ester. The desired filler can be obtained by a method or a method in which an optically active carboxylic acid is converted into an acid chloride and reacted in the presence of a detM N agent.

(2) A method in which a carboxylic acid derivative obtained by reacting an optically active carboxylic acid with an aminoalkylsilane is grafted onto an inorganic carrier having a hydroxyl group on its surface.

More specifically, N
, A method of dehydration condensation using a condensation reagent such as hexylcarbodiimide and 1-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline, and a method of converting carboxylic acid into an active ester and then condensing it with aminoalkylsilane. The objective can be achieved by grafting an organosilane represented by the general formula [I] obtained by a method or a method in which a carboxylic acid is converted into an acid chloride and reacted with an aminoalkylsilane in the presence of a dehydrochlorination agent onto an inorganic carrier such as silica gel. of filler is obtained.

The packing material having an optically active carboxylic acid 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, it is possible to
Separation and analysis of enantiomeric mixtures of compounds having - group, -0CO- group, -○H group, -0CONH- group or -NC6NH- group can be performed with good resolution and in a short time.

Hereinafter, the present invention will be explained in more detail with reference to Examples.

Example 1 Silica gel (average particle size 10 μm, average pore size 60 A1 surface area 500 doors/y) was dried in a vacuum drying mold at 1300 °C for 4 hours, and 8-aminopropyltriethoxysilane 20f was dissolved in 200 Me dehydrated toluene. and stirred at 60°C for 6 hours. The reaction product was filtered, the residue was washed 8 times with 100 rn1'' of acetone, and dried under reduced pressure to obtain 3-aminopropylsilylated silica gel (hereinafter referred to as AP).
The elemental analysis value of this product is N:1.
22%, C: 8.52%, which means that the 8-aminopropyl group is about 0.9% for the grafted silica gel 1y.
0mmol This corresponds to being crafted.

Separately, (-1-1-2-(4-chlorophenyl)isovaleric acid 3y was dehydrated by adding dimethylformamide 2m)
- Add 3Q me of hebutane to dissolve it, and add 21 thionyl chloride one by one over a period of about 20 minutes while stirring. After gently stirring at room temperature for 6 hours, the solvent was distilled off at room temperature under reduced pressure to obtain ((1)-2-(4-chlorophenyl)
Isovaleric acid chloride is obtained.

Next, take the entire amount of this (+l-2-(4-chlorophenyl)isovaleric acid chloride) and add the AP S 2.5 y
Suspended in 30 parts of dehydrated tetrahydrofuran, added to the fully degassed solution under reduced pressure, added 22 parts of trietheramine, washed with 5 methanol and water at room temperature, further washed with methanol and ethyl ether, and dried under reduced pressure. and ((1
The desired filler grafted with )-2-(4-chlorophenyl)isovaleric acid was obtained. The elemental analysis value of this is N
: 1.20%, C: 10, 56%, which indicates that about 0.68 mmol of (+l-2-(4-chlorophenyl)isovaleric acid was added to the grafted silica gel 12).

The filler thus obtained had an inner diameter of 4 mm.

Fill the slurry into a 25 cm long stainless steel column,
Under the following conditions (±)-〇-methylphenylacetic acid a, 5
- Dinitroanilide was analyzed and the chromatogram shown in Figure 1 was obtained.

Temperature: Room Temperature transfer Phase: n-hexane/1,2-dichloroethane/ethanol (100:20:l) Flow rate: 1 mc/min Detector: Ultraviolet absorption meter (wavelength: 254 nm) Peak in Figure 1 Number +1+ is (-)-α-methylphenylacetic acid 3.5-dinitroanilide, (2) is (-F)-α-
These are the peaks of methylphenylacetic acid 3,5-dinitroanilide.

The time required for the peak (2) to elute was approximately 13 minutes, the separation coefficient was 1.29, and the area ratio between the peaks of ftl and (2) was 50:50.

Example 2 (+)-) Lance-thin acid chloride 3y was taken and Example 1
The AP S 2.5 y obtained in step 2 was suspended in dehydrated tetrahydrofuran 3 Q me, added to the sufficiently degassed liquid under reduced pressure, triethylamine 2 y was added, and the mixture was slowly stirred at room temperature for 5 hours and then at 50°C for 8 hours. Stir. After cooling to room temperature, washing with tetrahydrofuran, methanol and then water, methanol and ethyl ether,
By drying under reduced pressure, the desired filler grafted with (deca-trans-dilute acid) was obtained.The elemental analysis value of this material was N:1.
20%, C: 11.27%, which means that (+)-trans-dilute acid is about 0.6% for the grafted silica gel 12.
Indicates that 7 mmol was grafted.

The filler obtained in this way has an inner diameter of 4 fl and a length of 25 mm.
The slurry was packed into a stainless steel column (±)-1-(4-chlorophenyl), -4,4-dimethyl-2-(1,2,4-triazol-1-yl)- under the following conditions.
1-Penten-3-ol was analyzed and the chromatogram shown in Figure 2 was obtained.

Temperature: Room temperature Mobile phase: n-hexane/isopropanol (24:1
) Flow rate: 1-7 minutes Detector: Ultraviolet absorption meter (wavelength: 254 nm) In Figure 2, peak number (1) is ←l-1-(4-chlorophenyl)-4,4-cy)f -(1,2,4-triazol-1-yl)-1-benzon-3-ol, (2)
)-1-(4-chlorophenyl)-4,4-if
Ru-2-(1,2,4-triazol-1-yl)-1
- Each peak of penten-3-ol.

The time required for peak (2) to elute is approximately 8 minutes.
The separation factor was 1.18, and the peak area ratio of (1) and (2) was 50:50.

Example 8 A filler (hereinafter referred to as CHR-5i) prepared by gelasting the (-1-1-2-(4-chloro(-1-)-) lance-thin acid obtained in Example 1 was used. omitted) each with an inner diameter of 4
The slurry was filled into a stainless steel column with a length of 25 mm, and the enantiomeric mixture of the following compounds was separated under the following conditions to determine the separation coefficient.

Temperature: Room Temperature Flow rate: 1 me/min Detector: Ultraviolet absorption meter (wavelength: 254nrn) The results are shown in the table below.

(Call, 1)” Two-note Ino 1 Compound Smell 7E, I'lN
B: 8,5-dinitrobenzoyl, Ac niacetyl, D
NA: 8,5-dinitroanilide.

r) NP: represents 3,5-dinitrophenyl.

(Note 2) In the above mobile phase, A: n-hexane/1,
2-dichloroethane/ethanol (1,00:20:
l), B: n-hexane/1,2-dichloroethane/ethanol (20:(i:1), C: n-hexane/isopropanol (199:1), D: n-hexane/isopropanol ( 24:1) 71°

[Brief explanation of the drawing]

Figures 1 and 2 show the chromadgrads obtained in Examples 1 and 2, respectively, where the vertical axis represents strength and the horizontal axis represents retention time. Figure-1 U b 70 15 (minutes) Figure-2 Procedural amendment (spontaneous October 27, 1980 Kazuo Wakasugi, Commissioner of the Patent Office, 1981 Patent Application No. 160518 2, Title of invention Optically active carboxylic acid Relationship to the Kuro incident where the patent applicant was grafted Address of patent applicant 5-15 Kitahama, Higashi-ku, Osaka Name (
209) Sumitomo Chemical Co., Ltd. Representative Hijikata
Take 4, Agent address: 5-15-5 Kitahama, Higashi-ku, Osaka, Clarification of the subject of amendment 1, Coverage of the scope of T+ patent Hrf sought and Detailed explanation of the invention column 6, Contents of correction (1) Patent The scope of claims will be revised as shown in the attached sheet. (2nd) Lines 11 to 10 from just below No. 7 of 11W in front of the gate are [-carboxylic acid residues, and B and 4 have one or more asymmetric carbon atoms." It represents a carboxylic acid residue. Correct ``Yo.'' (3) From page 7, line 9, and page 8, line 8, between "indicated" and "carboxylic acid derivative," respectively. Insert "optically active". Claims (1) General formula CI) I R2-8i-(OHz)n-Nu-CO-R4[:[:
]几3 [In the formula, R1, R2 and R3 are the same or different,
It represents an alkyl group, an alkoxyl group, a hydroxyl group, or a halogen atom, and at least one of R1, R2, and R3 is an alkoxyl group, and the alkoxyl group is a halogen atom. n is an integer from 2 to 4, and 4-CO represents various optically active carboxylic acid residues. ] A chromatographic packing material in which an optically active carboxylic acid derivative represented by the following formula is grafted onto an inorganic carrier having a hydroxyl group 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 CI), the carboxylic acid residue is an optically active a-arylalkylcarboxylic acid residue or a cyclopropanecarboxylic acid residue, Claim 1
Chromatographic packing material according to item 1 or item 2. (4) In the above general formula [■], the carboxylic acid residue is an optically active 2-(4-chlorophenyl)isovaleric acid residue, 2
-Phenylpropionic acid residue or -folic acid residue, Claims 1, 2 or I- is the chromatographic packing material according to Claim 3. (5) In the above general formula [■] , wherein the aminoalkylsilane residue is an OJ-aminopropyltriethoxysilane residue or an ω-aminopropyltrichlorosilane residue, Chromatographic media as described. (6) General formula [l] 几1 几2-8i-(CHz)n-NH-00-R4[11]
3 [wherein R1, R2 and R3 are the same or different,
represents an alkyl group, an alkoxyl group, a hydroxyl group, or a halogen atom; at least one of Rx, IL2, and 几3 is an alkoxyl group or a halogen atom; n is an integer from 2 to 4; 1. R4-
C0 represents various optically active carboxylic acid residues. ] An optically active carboxylic acid derivative represented by is grafted onto an inorganic carrier having a hydroxyl group on its surface. Using a chromatographic packing material, a 10 0 N H- group, -0CO, bonded to an asymmetric carbon. - group, -〇H group, -QC!
A liquid chromagraph analysis method characterized by separating (17) and analyzing an enantiomeric mixture of a compound having a 0NH- group and a -NOQNH- group. (7) The analytical method according to claim 6, which uses a chromatographic packing material in which the inorganic carrier having hydroxyl groups on its surface is silica gel. (8) Claim 6 using a chromatographic packing material in which the carboxylic acid residue/group is an optically active α-arylalkylcarboxylic acid residue or cyclopropanecarboxylic acid residue in the above general formula [I]. or the analytical method described in Section 7. (9) In the above general formula [■], the carboxylic acid residue is an optically active 2-(4-chlorophenyl)isovaleric acid residue,
Claim 6 uses a chromatographic packing material having 2-phenylpropionic acid residues or 2-phenylpropionic acid residues.
Chromatography in which the aminoalkylsilane residue in the above general formula [■] is an ω-aminopropyltriethoxysilane residue or an ω-aminopropyltrichlorosilane residue. An analytical method according to claim 6, 7, 8 or 9 using a graph filler.

Claims (1)

[Claims] (1) General formula [11 I] [wherein R1, R2 and R3 are the same or different,
It represents an alkyl group, an alkoxyl group, a hydroxyl group, or a halogen atom, and at least one of R1+R2 and R3 is an alkoxyl group or a halogen atom. n is an integer from 2 to 4, R4-C0 is various carboxylic acid residue, and R4 has one or more asymmetric carbon atoms. ] A chromatographic packing material in which a carboxylic acid derivative shown in Te is grafted onto an inorganic support 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 [11], the carboxylic acid residue is an optically active α-arylalkylcarboxylic acid residue or a cyclopropanecarboxylic acid residue, Claim 1
Chromatographic packing material according to item 1 or item 2. . (41 In the above general formula [1], the carboxylic acid residue is an optically active 2-(4-chlorophenyl)isovaleric acid residue, 2
The chromatographic packing material according to claim 1, 2 or 3, which is a phenylpropionic acid residue or a chrysanthemum acid residue. (5) Claims 1 and 2, wherein in the above general formula [I], the aminoalkylsilane residue is an ω-aminopropyl, triethoxysilane residue or ω-aminopropyltrichlorosilane residue, Chromatographic packing material according to item 8 or 4. +6j -・General formula [1,1 1 噸R2-8i -(CHz)1-NH-CO-Ra
IT]3 [wherein Ru, R2 and islands are the same or different and represent an alkyl group, an alkoxyl group, a hydroxyl group or a halogen atom, and at least one of R1+R2 and R3 is an alkoxyl group or a halogen atom . n is an integer from 2 to 4, and -CO is a variety of carboxylic acid residues, and the islands have one or more asymmetric carbons. A carboxylic acid derivative represented by group, -0C
A liquid chromatography analysis method characterized by separating and analyzing an enantiomeric mixture of a compound having an ONH- group and a -NCONH- group. (7) The analytical method according to claim 6, which uses a chromatographic packing material in which the inorganic carrier having hydroxyl groups on its surface is silica gel. (8) In the above general Nisan, the carboxylic acid residue is an optically active α-arylalkylcarboxylic acid! 5 units (J
The analytical method according to claim 6 or 7 using a chromatographic packing material which is a pancarboxylic acid residue (1) (9) In the general formula [Mon], a carboxylic acid residue or optically active 2-(4-chlorophenyl)isovaleric acid group,
Claim 6 uses a chromatographic packing material that is a 2-phenylpropionic acid residue or a tertiary chrysanthemum acid residue The sixth group is an ω-aminopropyltriethoxysilane residue or
- Claims 6 and 7 using a chromatographic packing material that is an aminopropyltrichlorosilane residue;
The analytical method according to item 8 or 9.