JPH06298672A - Chirality-discrimination agent and separating agent for chromatography - Google Patents

Abstract

PURPOSE:To provide a chirality-discrimination agent having a structure containing an organosilane compound grafted to an inorganic carrier having specific group on the surface, containing a ligand exchanging function part as well as hydrogen-bond interacting part and useful for the separation of optical isomer mixture. CONSTITUTION:The objective discrimination agent has a structure containing an organosilane compound expressed by formula (R1, R2 and R3 are alkyl, alkoxy, etc.; at least one of them is alkoxy or halogen; R4 is lower alkylene; R5 is residue produced by removing one amino group and one carboxyl group from an optically active amino acid or its D isomer; R6 is H, lower alkyl, etc.; the tartaric acid part and the amino acid or imino acid part have optically active forms) (e.g. aminopropyltriethoxysilane bonded with L-tartaric acid-valine) and grafted to an inorganic carrier (preferably a silicon-containing carrier such as silica gel) having hydroxyl group on the surface. A filler for chromatography can be prepared by using a proper grafting method.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to the use of the tartaric acid monoamide compound represented by the above general formula. For the above-mentioned applications, a ligand exchange functional unit capable of directly separating optical isomers such as amino acids and oxyacids and a hydrogen bond interaction site capable of separating derivatives of optical isomers such as amines, amino acids and carboxylic acids are combined. It has uses as a chiral discriminating agent and as a chromatographic packing material.

[0002]

As an optical resolution method by liquid chromatography utilizing tartaric acid, mono-n-octylamide of tartaric acid [(1) Journal of Liquid Chromatography (J.
Liq. Chromatogr. ) Vol. 9, p. 551 (1986)] or di-isopropylamide derivative [(2) Proceedings of the 46th Analytical Science Symposium, 53 (1985)] as a liquid phase separating agent. However, since these use a separating agent for the mobile phase, it is necessary to re-separate the separating agent from the eluent, which is complicated.

As a separating agent, (R, R) -tartaric acid-mono-isopropylamide (3), (R, R) -tartaric acid-mono-1-
(α-Naphthyl) ethylamide [(4) Japan Society for Analytical Chemistry 3rd
Proceedings of the 4th Annual Meeting, p. 310 (1985)] and (R,
R) -tartaric acid-mono-isopropylamide [(5) No. 48
Some of them are covalently bonded to silica gel to form a stationary phase, but the optical resolution is low because they are not of the ligand exchange type. Other than that, there is a method of covalently bonding (R, R) tartaric acid to silica gel to form a stationary phase [(6) Fresenius Zeitschrift Fuhr Analyticshemy (F
resenius Z. Anal. Chem.) Volume 320, 51-54
Page], this has a drawback that the optical resolution in the normal phase system is extremely poor because it does not have an amino acid amide moiety having a hydrogen bond interaction function effective for chiral discrimination in the normal phase system. The present invention seeks to provide a separating agent which does not have the above-mentioned drawbacks.

[0004]

[Means for Solving the Problems]

 General formula:

[Chemical 2] [Wherein R ₁ , R ₂ and R ₃ are the same or different,
It represents an alkyl group, an alkoxy group, a hydroxy group or a halogen atom, at least one of which is an alkoxy group or a halogen atom. R ₄ is a lower alkylene group, R ₅ is a residue obtained by removing one amino group and one carboxy group from an optically active amino acid or its D isomer, and R ₆ is hydrogen or a lower alkyl group, or R ₆ is a residue in which 1 imino group and 1 carboxy group are removed from imino acid or its D isomer together with R ₅ (provided that in R ₅ above, the remaining OH, NH, NH ₂ may be substituted with a group -Y-Z-, wherein -Y- is -CO- or -CONH-, and Z is lower alkyl, a monocyclic or bicyclic aryl group or a monocyclic or bicyclic aralkyl group. Is). The tartaric acid moiety and the amino acid or imino acid moiety are optically active forms. ] The organosilane compound represented by the formula [1] provides a chirality discriminating agent and a packing material for chromatography, which has a structure in which it is grafted onto an inorganic carrier having a hydroxy group on the surface.

In the above general formula (I), the alkyl group of R ₁ , R ₂ and R ₃ preferably has 1 to 20 carbon atoms, more preferably 1 to 12 carbon atoms, and 1-
6 is most preferable. Preferred are lower alkyl groups, including those having 1-6 carbon atoms.
The alkyl group may be linear or branched. Representative examples of alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, t-butyl, pentyl, hexyl and the like. R ₁ , R ₂ and R ₃
May be the same or different. The alkoxy group includes a group formed by the above alkyl group bonded to an oxygen atom. Preferred is a lower alkoxy group. The halogen atom includes chlorine, bromine, iodine and the like.

In the above general formula (I), the lower alkylene group for R ₄ is a linear or branched group having 1 to 6 carbon atoms, such as methylene, ethylene, trimethylene, 1
-Methylethylene, 2-methyltrimethylene, tetramethylene, pentamethylene, hexamethylene and the like are included.

The lower alkyl group for R ₆ is most preferably one having 1 to 6 carbon atoms. Representative lower alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl and the like.

The monocyclic or bicyclic aryl group includes a group having one or two benzene nuclei, for example, phenyl, tolyl, xylyl, cumeryl, naphthyl or methylnaphthyl. As the monocyclic or bicyclic aralkyl group, similarly, benzyl, phenethyl, naphthylmethyl, 1
-Includes naphthylethyl and the like. These may be substituted with a nitro group. When a nitro group is present on the ring of a monocyclic or bicyclic aryl group or a monocyclic or bicyclic aralkyl group, this is usually 1-5, preferably 1-3.
One, for example one or two, may be attached at any substitutable position. Specifically, for example, 3,5-dinitrophenyl, 3-nitrobenzyl and the like are included.

Asymmetric carbon atoms may be contained in R ₄ and R ₆ . As the amino acid, asparagine, aspartic acid, aminobutyric acid, alanine, β-alanine, arginine, isoleucine, ornithine, glutamine, glutamic acid, serine, tyrosine, tryptophan, threonine, norleucine, valine, histidine, phenylglycine, phenylalanine, methionine, lysine. , Leucine and the like, and imino acids include proline, hydroxyproline, histidine and the like. Many of these are naturally occurring and their homologues. These are usually derived from a carboxy group and an amino (imino) group bonded to the α carbon atom, but in some cases, they are derived from a group bonded to the ω carbon atom (eg, lysine).
The tartaric acid moiety is an optically active form, that is, (R, R) -tartaric acid [d-or _L- or _{R (R} *, _R * _)- (natural form) tartaric acid] form, and (S, S) tartaric acid [ l-or _D -or
_{S (R} *, _R * _)- tartaric acid] form, or either, is the predominant mixture. Also, the amino acid or imino acid moieties are in the optically active form, ie, the L or D form.

In the present invention, the inorganic carrier having a hydroxy group on the surface is preferably a silicon-containing carrier such as silica gel or porous glass, and the carrier may have any shape such as spherical or crushed. In order to obtain a high-yield chromatographic column, fine particles having a uniform particle size as much as possible are preferable. “Grafted structure” means that the actual production method is one-step grafting or multi-step grafting as long as it has a structure defined as a graft product of two parts as a result. It is meant to be included regardless of the reaction sequence in the case of a certain number of stages. During the grafting, some or all (usually 1 to 2) of the groups R _{1 to} R ₃ of the compound of formula I react with the hydroxy groups of the carrier to form an ether type bond —O—S.
form i.

In preparing the packing material for chromatography represented by the above formula (I), various grafting methods can be adopted. Examples of such grafting method include the following methods. An aminoalkylsilane is reacted with an inorganic carrier having a hydroxy group on the surface to introduce an aminoalkylsilyl residue on the surface of the inorganic carrier, and an optically active amino acid or imino acid and tartaric acid are introduced into the residue.
A method of reacting (functional group may be protected appropriately) and dehydration condensation.

Specifically, the inorganic carrier having a hydroxy group on the surface is represented by the general formula (II)

[Chemical 3] [Wherein R ₁ , R ₂ , R ₃ and R ₄ have the same meanings as described above. ] The aminoalkylsilane represented by the formula] is reacted to introduce an aminoalkylsilyl residue onto the surface of the inorganic carrier, and then the general formula

[Chemical 4] And a reactive derivative at the carboxy group represented by

[Chemical 5] The desired filler can be obtained by sequentially reacting the carboxylic acid represented by or the reactive derivative at the carboxy group and dehydration condensation. In the above formula, P ₁ , P ₂ , P
₃ and P ₄ are protecting groups commonly used in peptide chemistry as protecting groups for amino, hydroxy or carboxy groups. Such protecting groups are described in "New Experimental Chemistry Course", Volume 14, together with their elimination methods.

Specifically, the "amino protecting group" is described, for example, in "New Experimental Chemistry Course", Vol. 14, pp. 2555-2569. For example, as an acyl type, formyl (HCl / CH ₃ OH), lower alkanoyl groups such as acetyl (elimination with acid or alkali), halo lower alkanoyl groups such as 2-chloropropionyl (elimination with acid or alkali), benzoyl (elimination with acid or alkali), etc. Arylcarbonyl group, an aryl lower alkanoyl group such as phenylpropionyl (elimination with acid or alkali),
Examples of the urethane type include lower alkoxycarbonyl groups such as methoxycarbonyl, ethoxycarbonyl, tertiary butoxycarbonyl (elimination with HBr or HCl / AcOH), benzyloxycarbonyl (H ₂ / Pd, HI or HBr or HCl / AcOH). Etc.), 2- (p-biphenylyl) isopropoxycarbonyl (AcOH / HCO
Aryl lower alkyloxycarbonyl group (dissociated with OH, trifluoroacetic acid, etc.), aralkyl type benzyl (dissociated with H ₂ / Pd, Na / NH ₃ ), trityl
Aryl lower alkyl groups such as (elimination with H ₂ / Pd) and azomethine type benzylidene (H ₂ / Pd,
Aryl lower alkylidene groups such as (elimination with HCl).

The "hydroxy protecting group" is described, for example, in "New Experimental Chemistry Course", Vol. 14, p. 2497-2516, and methyl (elimination by hydrolysis after oxidation), tertiary butyl.
(Lowering with CF ₃ CO ₂ H), lower alkyl group such as benzyl (eliminating with H ₂ / Pd, NH ₃ / Na), trityl (eliminating with acetic acid), trimethylsilyl (C ₂ H ₃ OH), tri-lower alkylsilyl groups such as tertiary butylmethylsilyl (elimination with acetic acid), aryl lower alkoxy lower alkyl groups such as benzyloxymethyl (elimination with NH ₃ / Na), ethoxycarbonyl (KOH /
Lower alkoxycarbonyl group such as elimination with CH ₃ OH),
Aryl lower alkoxycarbonyl group such as benzyloxycarbonyl (elimination with H ₂ O ₂ / CH ₃ CO ₂ H), acetyl (elimination with acid or base), formyl (KHCO ₃ / H ₂
Lower alkanoyl group such as O / CH ₃ OH), nitric acid (elimination with Na ₂ S), sulfonic acid (Na / Hg / CH ₃ O)
Acid residue such as elimination with H), methylene acetal (CH ₃ C
Lower alkylidene acetal groups such as O ₂ H- (CO ₃ CO ₂ ) ₂ O-H ₂ SO ₄ ) and ethylidene acetal (elimination with CH ₃ COOH), aryls such as benzylidene acetal (elimination with acid) Examples thereof include acetal group.

The "carboxy protecting group" is described, for example, in "New Experimental Chemistry Course", Vol. 14, pp. 2535-2544, and includes, for example, methyl (elimination with acid or alkali), ethyl (same as above), third. Butyl (HCl / CH ₂ Cl ₂ , CF ₃ CO
OH, TsOH / AcOH, etc.) and other lower alkyl groups such as 2,2,2-trichloroethyl (Zn / AcOH, HC
Halo lower alkyl group such as OOH, etc.), lower alkylthio lower alkyl group such as methylthioethyl (CH ₃ I and then alkali treatment), methoxymethyl (elimination with acid)
Lower alkoxy lower alkyl groups such as tosylethyl (Na
Arylsulfonyl lower alkyl group such as OH / dioxane), optionally substituted arylthio lower alkyl group such as p-nitrophenylthioethyl (oxidized to sulfone and then eliminated with alkali), benzyl (H ₂ / Pd , Na /
NH _3, NaOH / dioxane, desorption with HBr / AcOH etc.), p-methoxybenzyl _{(H 2 / Pd, CF 3} COO
H, HCOOH, etc.), 2,4,6-trimethoxybenzyl (HBr / AcOH, elimination), pentamethylbenzyl
(Elimination with CF ₃ COOH), p-nitrobenzyl (H ₂ / P
d, desorbed with NaOH), benzhydryl (H ₂ / Pd, NaO
H)), trityl (NaOH, HCI / CH ₃ OH), anthranylmethyl (HBr / AcOH, NaOH,
And optionally substituted lower arylalkyl groups such as CF ₃ COOH) and hydroxamic acid (elimination with HIO ₄ ). The terms "carboxy protecting group", "hydroxy protecting group" and "amino protecting group" above have the same meaning as those terms used in peptide synthesis.

The aminoalkylsilane is preferably ω-aminoalkylalkoxysilane or ω-aminoalkylhalogenosilane, and examples thereof include ω-aminopropyltriethoxysilane and ω-aminopropyltrichlorosilane. In the above-mentioned production method, the reaction of introducing the organosilane to the surface of the inorganic carrier can be carried out according to a conventional method commonly used for silylation of a compound having a hydroxy group. This reaction is generally performed in an organic solvent at a relatively mild temperature. The reaction between any carboxylic acid or a reactive derivative at the carboxy group thereof and an inorganic carrier having an aminoalkylsilyl group introduced thereinto or the above-mentioned carrier having an amino acid moiety bound thereto can be carried out by, for example, amide bond formation reaction commonly used in peptide synthesis. It can be performed according to a conventional method. As the reactive derivative, an acid halide, an acid anhydride (for example, an alkylcarbonic acid mixed acid inorganic substance), an active ester or an active amide (for example, imidazolide) is used.When the acid itself is used, the reaction is performed in the presence of a condensing agent. It is advantageous to do so. As the condensing agent, carbodiimides, carbonyldiimidazoles,
Woodward reagent, N-alkoxycarbonyl-2-alkoxy-1,2-dihydroquinoline and the like are used.
The reaction conditions vary depending on the combination of the compounds to be reacted, and the reaction is carried out in an organic solvent or an aqueous solvent under cooling or at room temperature or under heating, and optionally in the presence of a base such as a tertiary organic amine. Removal of the protecting group is carried out by a conventional method.

The packing material obtained by the present invention can be packed in a column for chromatography according to a conventional method and used as a stationary phase for optical resolution of a mixture of optical isomers by liquid chromatography. That is, it is packed in a column of glass, stainless steel, titanium or the like having an appropriate diameter and length, and a solution containing a mixture of optical isomers is back-injected from the inlet side, and an appropriate solvent and solvent conditions, particularly commonly used By developing and fractionating under conditions of normal phase partition or reverse phase partition, various optical isomer mixtures can be separated and analyzed.

[0018]

EFFECTS OF THE INVENTION Since the compound (I) of the present invention has a remarkable difference in interaction with optical isomers, this property can be utilized as a chirality discriminating action. Therefore, the filler of the present invention has excellent chirality discrimination.
Thus, the packing material of the present invention is capable of chromatographically separating different optical isomer mixtures, for example different esters, alcohols, carboxylic acids, amines and amino alcohols and their derivatives. It falls into the broadest category of fillers. In particular, as the chiral recognition action for separating optical isomers, there are various interactions between the chiral moiety of the stationary phase and the sample, but conventionally, the hydrogen bonding interaction and the reverse phase which are effective in the positive phase system are exhibited. There is no stationary phase that also has a ligand exchange interaction that exerts an effect in a system (water system), and the stationary phase of the present application is excellent in that it has both of them, that is, an amine, a carboxylic acid, an alcohol in a normal phase system. The optical isomers of can be separated, and the optical isomers of amino acids and oxyacids can be directly separated in a reverse phase system (water system), and therefore a very wide range of compounds can be optically resolved.
Since the types of eluents that can be used are among the broadest in the existing stationary phases (there are almost no restrictions other than high-concentration acids and bases), there is a wide range of mobile phase modifications for successful resolution. It is possible to try.

Moreover, the filler of the present invention has the advantages that it is easy to manufacture because it has a small number of manufacturing steps, and that it is also excellent in durability because the underlying compound is chemically stable. In addition, a high number of theoretical plates can be obtained as a column, and there is little variation in quality. Hereinafter, the present invention will be described with reference to examples, but the examples do not limit the present invention.

[0020]

【Example】

Example 1: Silica gel (average particle size 5 μm, average pore size 120
Å, 200 g of surface area 330 m ² / g) was dried under reduced pressure at 120 ° C. for 2 hours, and then 200 g of 3-aminopropyltriethoxysilane was added to a solution of 1 liter of dehydrated detrahydrofuran and heated under reflux for 3 hours. The solution was left to stand, the gel was sufficiently sedimented, and the supernatant was discarded. Methanol was added to this and stirred, and the supernatant was similarly decanted. Furthermore, the residue is washed with tetrahydrofuran, chloroform / methanol mixture, chloroform and hexane (500 ml each), dried,
3-Aminopropylsilylated silica gel (hereinafter abbreviated as APS) was obtained. The elemental analysis value of this product is N: 1.32
%, C: 4.78%, which corresponds to about 0.94 mmol of 3-aminopropylsilyl group grafted to 1 g of this product.

Then, a solution prepared by dissolving 8.2 g of t-butoxycarbonyl-L-valine in 180 ml of dehydrated tetrahydrofuran was ice-cooled, and 1-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) 10. After adding 8 g and stirring with ice cooling for another 1 hour, 15 g of the above APS was added and suspended, and the mixture was stirred overnight at room temperature. The reaction product was filtered, and the residue was washed twice with a mixture of chloroform / methanol, 180 ml each of methanol, chloroform and hexane and dried, and 16.1 g of silica gel grafted with t-butoxycarbonyl-L-valylaminopropylsilyl group. Got The elemental analysis value of this product is N: 1.92%, C: 1.
It was 1.0%. Then, 16.0 g of this gel is added to 50%
180 ml of a trifluoroacetic acid dichloromethane solution was added and suspended, and the mixture was stirred at room temperature for 1 hour. The reaction was filtered, the residue washed with 180 ml of dichloromethane and then stirred in ammonia-dichloromethane solution for 20 minutes. The reaction product was filtered, and the residue was washed twice with 180 ml each of methanol, dichloromethane and hexane and dried. From the above gel, 14.9 g of grafted silica gel in which the t-butoxycarbonyl group was eliminated was obtained. The elemental analysis values of this product were N: 2.03% and C: 7.97%.

Further, 10.6 g of this gel was added to dehydrated tetrahydrofuran in which 11.5 g of diacetyl-L-tartaric anhydride was dissolved, and the suspension was heated and refluxed for 4 hours and a half. After cooling, the reaction product was filtered, and the residue was washed twice with 100 ml each of methanol, chloroform and hexane and dried to have the same configuration as diacetyl-L-tartaric acid.
-Carboxy-2,3-diacetoxypropionyl-L
11.6 g of silica gel grafted with valylaminopropylsilyl groups are obtained. The elemental analysis value of this product is N:
It was 1.83% and C: 12.4%.

In order to block the residual amino groups of ASP and valine, 11.5 g of the above gel was added to 100 ml of dehydrated tetrahydrofuran in which 2.4 g of acetic anhydride was dissolved, and the mixture was heated under reflux for 3 hours. The reaction product was filtered, and the residue was washed twice with 100 ml each of methanol, chloroform and hexane and dried to obtain 11.1 g of gel. The elemental analysis values of this product were N: 1.72% and C: 12.2%.

Finally, 6.0 g of this gel was suspended in 300 ml of an aqueous solution of perchloric acid having a pH of 3.0 and containing 5% of acetonitrile, and the suspension was heated and refluxed for 8 hours. The mixture was left standing to cool, the gel was allowed to settle sufficiently, the supernatant was decanted, and 150 ml of methanol was added and stirred. After allowing to stand, the supernatant was similarly decanted, the residue was washed twice with 150 ml each of chloroform and hexane, dried, and 3-carboxy-2,3-dihydroxy having the same configuration as L-tartaric acid. 5.0 g of the desired filler having grafted propionyl-L-valylaminopropylsilyl group was obtained. The elemental analysis value of this product is N: 1.37
%, C: 7.55%. This indicates that about 0.49 mmol of L-tartaric acid-L-valine was grafted on 1 g of this product. The packing material thus obtained was slurry-filled into a stainless steel column having an inner diameter of 4 mm and a length of 25 cm. Using the column adjusted as above,
Organic solvent system (normal phase system) and copper ion aqueous solution system (ligand exchange system)
The racemates of the various mixtures were separated by two different eluents of. An example of the result of the organic type is shown in Table 1 and an example of the result of the copper type is shown in Table 2. Temperature: room temperature Eluent: shown in Tables 1 and 2. Eluent flow rate: 1.0 ml / min Detector: UV absorber (wavelength 254 nm or 230 n
m)

[Table 1]

[Table 2] In this table, the retention factors (K ₁ 'and K ₂ ') and separation factor (α) are calculated by the following equation.

[Equation 1] t ₁ : Retention time of enantiomer that is weaker adsorbed t ₂ : Retention time of enantiomer that is more strongly adsorbed t ₀ : Retention time corresponding to dead volume of column Separation coefficient is completely optical when α = 1 It shows that there is no resolution and that the optical resolution increases as the difference from 1 increases.

Example 2 In Example 1, t-butoxycarbonyl-D-phenylglycine was used in place of t-butoxycarbonyl-L-valine, and the same operation was carried out.
A silica gel grafted with 3-carboxy-2,3-diacetoxypropionyl-D-phenylglycylaminopropylsilyl group having the same configuration as diacetyl-L-tartaric acid was obtained. This was added to 20% chloroform trifluoroacetate, suspended, and stirred at room temperature for 3 hours. The reaction product was filtered, and the residue was washed twice with chloroform, methanol, chloroform and hexane. Dried
3-carboxy- having the same configuration as L-tartaric acid
A desired filler obtained by grafting 2,3-dihydroxypropionyl-D-phenylglycylaminopropylsilyl group was obtained. The elemental analysis value of this product is N: 1.43%,
C: 10.5%, which indicates that 0.5 g of L-tartrate-D-phenylglycine was grafted to 1 g of this product. The packing material thus obtained was slurry-filled into a stainless steel column having an inner diameter of 4 mm and a length of 25 cm. Using the column prepared as described above, racemates of various compounds were separated by two kinds of eluents having different properties: an organic solvent system (normal phase system) and a copper ion system (ligand exchange system). Table 3 shows an example of the results for organic systems.
Table 4 shows an example of the results for the copper system. Temperature: room temperature Eluent: Tables 3 and 4 Eluent flow rate: 1.0 ml / min Detector: UV absorber (wavelength 254 nm or 230 n
m)

[Table 3]

[Table 4]

Example 3: N- in 90 ml of acetonitrile
[N'-R-1- (α-naphthyl) ethylcarbamoyl]
Dissolve 9.4 g of succinimidate and add N ⁶ -t-
45 ml of an aqueous solution containing 7.4 g of butoxycarbonyl-L-lysine and 3.0 g of triethylamine was added, and the mixture was heated under reflux for 1 hour. After cooling, the residue was removed by filtration, 450 ml of water was added to dilute, the mixture was acidified with citric acid and extracted with chloroform. The solution was dried and concentrated to give N ² -R-1- (α- naphthyl) ethylcarbamoyl -N ^{6-t-butoxycarbonyl} -L- lysine 11.8 g.

In Example 2, the compound of the above-mentioned synthesis was used in place of t-butoxycarbonyl-D-phenylglycine, and similarly, N ^6- (3-carboxy-2, 3-dihydroxy-propionyl) -N ²
-[R-1- (α-naphthyl) ethylcarbamoyl] -L
A desired filler having a lysylaminopropylsilyl group grafted was obtained. The elemental analysis value of this product is N: 1.82
%, C: 10.9%, which means that N ^6- (L-tartaric acid) -N ^2- [R-1- (α-naphthyl) ethylcarbamoyl] -L-lysine was added to 1 g of this product. It shows that about 0.33 mmol was grafted. The packing material thus obtained was slurry-filled into a stainless steel column having an inner diameter of 4 mm and a length of 25 cm. Using the column prepared as described above, racemates of various compounds were separated by two different eluents, an organic solvent system (normal phase system) and a copper ion aqueous solution system (ligand exchange system). . Fifth example of organic results
Table 6 shows an example of the results for the copper system. Temperature: room temperature Eluent: shown in Tables 5 and 6. Eluent flow rate: 1.0 ml / min Detector: UV absorber (wavelength 254 nm or 230 n
m)

[Table 5]

[Table 6]

[Brief description of drawings]

FIG. 1 N-3,5-dinitrobenzoyl- (R, S)
It is a graph which shows an elution curve when -1-phenylethylamine is divided using the separating agent of the present invention.

FIG. 2 is a graph showing an elution curve when D, L-naphthylglycine was separated using the separating agent of the present invention.

[Explanation of Codes] 1 ... Peak of N-3,5-dinitrobenzoyl- (S) -1-phenylethylamine. 2 ... N-3,5-dinitrobenzoyl- (R) -1-phenylethylamine peak. 3 ... Peak of D-naphthylglycine. 4 ... L-naphthylglycine peak.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Yasuhiro Matsushita 3-1-1135 Kasugade, Konohana-ku, Osaka City, Osaka Prefecture Sumika Chemical Analysis Service, Ltd. (72) Inventor Naoko Kisu Kasuga, Konohana-ku, Osaka City, Osaka Prefecture Ichinaka 3-1-135 Sumika Chemical Analysis Service, Ltd.

Claims (2)

[Claims] 1. A general formula: [Wherein R ₁ , R ₂ and R ₃ are the same or different,
It represents an alkyl group, an alkoxy group, a hydroxy group or a halogen atom, at least one of which is an alkoxy group or a halogen atom. R ₄ is a lower alkylene group, R ₅ is a residue obtained by removing one amino group and one carboxy group from an optically active amino acid or its D isomer, and R ₆ is hydrogen or a lower alkyl group, or R ₆ is a residue in which 1 imino group and 1 carboxy group are removed from imino acid or its D isomer together with R ₅ (provided that in R ₅ above, the remaining OH, NH, NH ₂ may be substituted with a group -Y-Z-, wherein -Y- is -CO- or -CONH-, and Z is lower alkyl, a monocyclic or bicyclic aryl group or a monocyclic or bicyclic aralkyl group. Is). The tartaric acid moiety and the amino acid or imino acid moiety are optically active forms. ] The chirality discriminating agent having a structure in which the organosilane compound represented by the formula [3] is grafted to an inorganic carrier having a hydroxy group on the surface. 2. A packing material for chromatography comprising the compound according to claim 1.