JPH0430932B2 - - Google Patents

Description

[Detailed description of the invention]

(Industrial Application Field) The present invention relates to chromatographic separation technology, and more particularly, to a chromatographic stationary phase capable of separating, purifying, and analyzing optical isomers from a mixture of chiral substances; Regarding its manufacturing method. (Prior art and problems to be solved by the invention) In chromatographic methods such as gas chromatography and liquid chromatography, mixtures of equal amounts (racemates) of optical isomers such as amino acids, derivatives, and hydroxy acids are used. , optically active stationary phases have been studied and put into practical use in order to optically resolve each optical isomer (for example, Hara and Tsuchihashi, "The Realm of Chemistry", Nankodo, Tokyo, 1981 Special Edition 132).
No. 171, and “Bunseki” co-authored by Tsuchibashi and Hara, volume 9.
633 pages 1981). This stationary phase is usually of a chemically bonded type that is chemically bonded to the surface of a substrate in order to prevent performance deterioration due to flow of the stationary phase during use, and is used as a packing material when a column is used. However, conventional optically active chemically bonded stationary phases have selectivity with respect to the target of separation, and therefore the types of optically active substances that can be separated with a certain stationary phase are very limited, and only When trying to separate and analyze substances, there is a problem in that a stationary phase that is compatible with each substance must be used. The present invention was made in view of the above-mentioned circumstances, and its purpose is to provide a stationary phase for chromatography that can separate and analyze many types of optically active substances and has a wide range of applications, as well as a method for producing the same. It is to provide. (Means for Solving the Problems) The present inventors have found that a chemically bonded stationary phase having a special residue is effective in achieving the object of the present invention, and have completed the present invention. That is, in the many years of experimental research on optical resolution by chromatography, the present inventors introduced chiral (R,R)-N,N'-diisopropyltartrate amide (DIPTA) into the mobile phase of silica gel chromatography. When used as an additive
Due to molecular association between DIPTA and solute through hydrogen bonds, β-oxycarbonyl compound, 1,2
Obtained the knowledge that diols and α-amino acid derivatives are optically resolved (Tetrahedron Lett., 25,
329, (1984)). As a result of further testing and research based on this knowledge, it was confirmed that carbamoylhydroxycarboxylic acid amide has an asymmetric discriminating power that allows it to optically resolve optically active substances. The present invention was completed by successfully chemically bonding the group to the substrate. Stationary Phase for Chromatography The chiral stationary phase for chromatography of the present invention is chemically bonded to a substrate characterized by having a chiral carbamoylhydroxycarboxylic acid amide residue represented by the following structural formula (I). It is something. (In the formula, R ₁ , R ₂ and R ₃ mean a hydrogen atom, an alkyl group or an aryl group, and n is an integer of 1 to 6.) As a preferred embodiment of the present invention, R ₁ in the above formula (I) , R ₂ and R ₃ can be a hydrogen atom, a straight or branched alkyl group having 1 to 6 carbon atoms, a phenyl group, a naphthyl group or an anthryl group. This suitable bulky substituent is expected to have an advantageous steric effect on the stationary phase of the present invention. In yet another embodiment, n in formula (I) is 2,4
Or it is preferable to set it as an even number of 6. This embodiment imparts axial symmetry to the carbamoylhydroxycarboxylic acid amide residue of formula (I), thereby imparting good asymmetric discriminating power to the stationary phase of the present invention. Next, the stationary phase of the present invention will be explained in more detail. Chromatography that uses the stationary phase of the present invention includes gas chromatography, liquid chromatography, thin layer chromatography, paper chromatography, column chromatography, etc., and is applied to various chromatography. can do. In this specification, the stationary phase refers to a phase that does not move with the sample, and is in contrast to the mobile phase.
The stationary phase of the present invention has a chiral carbamoylhydroxycarboxylic acid amide residue represented by the above structural formula (I), and is chemically bonded to a substrate. The residue of formula (I) is preferably present in the skin portion of the stationary phase since this residue portion exhibits asymmetric discriminating power. The stationary phase of the invention is chemically bonded to a substrate. This type of chemical bond is
Any bond may be used as long as it does not contradict the purpose of the present invention, such as a covalent bond, an ionic bond, a coordinate bond, a hydrogen bond, etc. In the stationary phase of the invention, the residue of formula (I) is bound to the substrate, either directly or via a spacer. In the present invention, the spacer may be any spacer as long as it connects the residue of formula (I) and the substrate surface at regular intervals, and for example, has 1 to 1 carbon atoms.
10 linear or branched aliphatic carbon chains, aromatic carbon chains such as phenylene groups, and heterocarbon chains having heteroatoms in the main chain or side chains. The substrate used in the present invention can be determined depending on the type of chromatography, the target of analysis, etc., and includes, for example, silica, silica gel, activated carbon, various porous polymers, zeolite, isolite,
These include diatomaceous earth, alumina, porous glass, semi-rigid gels such as polystyrene gel, solids such as cellulose and polyamide, and polymeric fluids containing hydroxyl groups such as polyethylene glycol and polyvinyl alcohol. The stationary phase of the present invention has a carbamoylhydroxycarboxylic acid amide residue represented by the above structural formula (I). This residue contains at least one asymmetric carbon and has chirality. Method for producing a stationary phase The method for preparing the starting material for the method for producing a stationary phase of the present invention will first be described as an example. The starting materials for this process can be prepared starting from chiral mono- or polyhydroxydicarboxylic acids. () The dicarboxylic acid is converted to an acid anhydride as shown in the reaction formula below, and then reacted with an amine or alcohol to convert it into a half-amide or half-ester. (R ₁₁ = alkyl group) () The dicarboxylic acid is converted into an acyloxydicarboxylic anhydride by protecting the hydroxyl group (e.g., acylation) and deriving it into an acid anhydride as shown in the reaction formula below. Converted to half-amide or half-ester by reaction with amine or alcohol. The above two equations are for X=OH, but X
When is a halogen, the carboxyl group of the half-amide or half-ester can be converted into an acid halide by known methods. Also,
Groups that activate carbonyl groups, such as succinimide and derivatives thereof, can be similarly prepared by known methods. Next, the manufacturing methods (A) to (F) of the stationary phase of the present invention will be specifically explained. (A) A chiral carbamoyl hydroxycarboxylic acid derivative of formula (a) is condensed with an amino group chemically bonded to a substrate represented by the following structural formula () to produce the stationary phase of the present invention as shown in the reaction formula below. (method according to claim 3). (In the formula, R ₄ means a carbon chain or direct bond connecting the adjacent nitrogen atom and the substrate, and Car means the substrate) If necessary, the derivative (a)
The hydroxyl group of is converted into an acyloxy group, preferably an acetoxy group (OAc), for example by an acylating agent.
It is possible to protect the compound as shown in the reaction formula below, and then deprotect it by known hydrolysis after condensation, as shown in the reaction formula below. In formulas (a) and (a'), X is a hydroxyl group (-
OH), a reagent (dehydration condensation agent) that activates the carbonyl (C=O) of the carboxyl group, such as dicyclohexylcarbodiimide (DCC), can be added to promote the condensation reaction. This method allows a chiral stationary phase to be covalently linked to a substrate. (B) Neutralizing the carboxyl group of the chiral hydroxycarboxylic acid derivative of formula (a) with the amino group bonded to the substrate of formula (V) to produce the stationary phase of the present invention as shown in the reaction formula below. (The method described in claim 5). (In the formula, R ₅ and R ₇ represent a hydrogen atom, an alkyl group or an aryl group.) By this method, a chiral stationary phase can be linked to a substrate via an ionic bond. The starting material (a) in this production method is the formula (a') or (
It can be prepared by condensing the compound of a) with an amino acid or by subsequent deprotection by hydrolysis. In this method, if necessary, the hydroxyl group of formula (a) may be protected as an acyloxy group using an acylating agent, for example, before neutralization, and deprotected by known hydrolysis after neutralization. can. (C) The stationary phase of the present invention can be produced by condensing the alkoxy group of the chiral trialkoxysilane derivative of formula (a) with the hydroxyl group of the substrate as shown in the following reaction formula (Claim 6). (method described in section). (In the formula, R ₈ , R ₉ and R ₁₀ mean an alkyl group.) In this method, as shown in the reaction formula below, the hydroxyl group of formula (a) is optionally added to an acylating agent, for example, before condensation. It can be protected as an acyloxy group by , and after condensation, it can be deprotected by known hydrolysis. The starting material (a') in this production method can be obtained by amidating the compound of the formula (a') to give an alkoxysilane derivative, as shown in the following reaction formulas () and (). Compound (a) whose hydroxyl group is not protected can also be prepared in a similar manner. (D) The chiral hydroxycarboxylic acid derivative of formula (b) is condensed with an amino group chemically bonded to the substrate represented by formula (), and then the alkoxycarbonyl group is replaced with a carbamoyl group, and the following reaction formula is obtained: The stationary phase of the present invention can be manufactured as follows (method according to claim 7). In this method, if necessary, the hydroxyl group is protected as an acyloxy group, preferably an acetoxy group, using an acylating agent before the condensation, and after the condensation, known hydration is performed. Deprotection can be achieved by decomposition, and an alkoxycarbonyl group can be substituted with a carbamoyl group. When X in formulas (b) and ('b) is a hydroxyl group, it is desirable to add a reagent that activates the carbonyl of the carboxyl group, such as dicyclohexylcarbodiimide (DCC), to promote condensation. (E) The carboxyl group of the chiral hydroxycarboxylic acid derivative of formula (b) is neutralized with the amino group bonded to the substrate of formula (V), and then the alkoxycarbonyl group is replaced with a carbamoyl group to form the reaction formula shown below. The stationary phase of the present invention can be produced as follows (method according to claim 9). The starting materials (b) and ('b) for this process can also be prepared and the hydroxyl groups protected as described in (B) above. (F) condensing the alkoxy group of the chiral trialkoxysilane derivative of formula ('b) with the hydroxyl group of the substrate, then substituting the alkoxycarbonyl group with a carbamoyl group, and deprotecting it by hydrolysis to return it to the hydroxyl group, The stationary phase of the present invention can be produced as shown in the reaction formula below (method according to claim 10). In this method, the stationary phase of the invention can be produced without protecting the hydroxyl groups. The starting material ('b) or (b) in this production method can be prepared in the same manner as explained in (C) above. Reaction conditions (solvent,
reaction temperature, reaction atmosphere, operation, reaction time, etc.)
It is desirable to decide appropriately depending on the reagents used, etc. (Function) The mechanism of optical resolution of an optical isomer mixture such as a racemate by the stationary phase for chromatography of the present invention is not necessarily theoretically clear, but it is thought to work as follows. Since the stationary phase of the present invention linked to the substrate by a chemical bond has a chiral carbonylhydroxycarboxylic acid amide residue represented by formula (I), there are hydroxyl groups and amide groups on the asymmetric carbon,
These functional groups interact with the target to be separated (e.g., an equal mixture of the d- and l-isomers) to form diastereomeric complexes with the enantiomers, and the differences in this interaction allow for a wide range of possibilities. It is thought that it exhibits asymmetric recognition ability for compounds. (Effects of the Invention) The stationary phase of the present invention can provide the following effects. (a) Compared to conventional stationary phases, the stationary phase of the present invention has a wider range of applicability, and in chromatography, diols, α- or β-hydroxy acids,
Optical resolution of α-amino acid esters, amide derivatives, etc. can be performed. (b) β-hydroxyketone, which was thought to be difficult to optically resolve using conventional stationary phases,
Optical resolution can be performed using the stationary phase of the present invention. (Examples) Hereinafter, the present invention will be explained in detail with reference to examples. Example 1 Preparation of starting materials Tartaric anhydride diacetate (a) [Reference, Organic
Synthesis, Col4, 242 (1963)] 3.866g of anhydrous
It was dissolved in 35 ml of CH ₂ Cl ₂ and 3.4 ml of isopropylamine was added dropwise to this solution at 0°C. After the reaction, the solvent was distilled off under reduced pressure, and 30 ml of water and an appropriate amount of concentrated hydrochloric acid were added to the residue to adjust the pH to less than 1. This solution was extracted with 500 ml of ethyl acetate, and the ethyl acetate layer was washed with saturated brine. After washing, the ethyl acetate layer was dried with anhydrous sodium sulfate and further concentrated under reduced pressure. 4.789g of this concentrate was mixed with isopropyl alcohol-n-
Recrystallization from hexane gave 4.38 g of colorless crystals of monoamide (b). ¹ of the obtained monoamide (b)
The analysis results of HNMR spectrum are shown below. ^1H -NMR ( _CDCl3 -DMSO-d6), δ: 1.44
(3H, d, J=6.3Hz), 1.48 (3H, d, J=6.3
Hz), 2.10 (3H, s), 2.17 (3H, s), 3.83−4.27
(1H, m), 5.57−5.68 (2H, m), 6.67 (1H, brd,
J=7.5Hz), 10 (1H, brs). Example 2 Preparation of covalent stationary phase Add 1.6 g of the monoamide (b) obtained in Example 1 and 1.21 g of aminopropyl silica (c) to 10 ml of anhydrous tetrahydrofuran (THF), and add 1 ml of anhydrous THF at 0°C.
An additional 2.4 g of dicyclohexylcarbodiimide (DCC) dissolved in water was added. The reaction mixture was warmed to room temperature, 100 ml of chloroform was added to the reaction mixture, and the mixture was centrifuged. After the separation treatment, the supernatant was removed and the silica derivative was collected by filtration, washed with chloroform, methanol, acetone, and n-hexane, and dried. As a result, 1.28 g of a colorless solid substance (d) was obtained. IR (KBr) spectrum analysis result: 1760cm ^-1 ,
1670cm ^-1 , 1550cm ^-1 . 2 ml of anhydrous methanol was added to 996.6 mg of the obtained diacetoxy derivative (d), and a solution of 1.4 g of ammonia absorbed in 10 ml of methanol was added at 0° C. for hydrolysis. Filter the slurry after the reaction,
After washing with methanol, chloroform, and n-hexane, the residue was dried to obtain 924 mg of hydroxy compound (e). IR (KBr) spectrum analysis result: 1650cm ^-1 ,
1540cm ^-1 . Elemental analysis: C; 8.00%, N; 1.85%, H; 1.45
% Example 3 Reaction to optical resolution The stationary phase for chromatography prepared in Example 2 was slurried packed into a column with a diameter of 1 mm and a length of 50 cm.
A β-hydroxycarboxylic acid amide represented by the following structural formula was isolated. This analysis was carried out using 4 as the solvent.
% 2-propanol-n-hexane at a temperature of 20°C and 60 μl/hexane using 230 nm ultraviolet light as a detector.
The flow rate was min. The analysis results are shown in Table 1.

【table】

[Table] In general, the practical ranges of the retention ratio k' and the separation coefficient α are 1 to 10 and 1.1 or more, respectively, so that the stationary phase of the present invention is suitable for various β-hydroxycarboxylic acid amides. It can be seen from Table 1 that the Example 4 Preparation of ion-bonded stationary phase 1.24 g of monoamide (b) obtained in Example 1 and 528.2 mg of N-hydroxysuccinimide were added to 20 ml of anhydrous THF, and 1.0 g of DCC was dissolved in 1 ml of anhydrous THF at 0°C.
g was added. The precipitated urea derivative was filtered off and washed with anhydrous THF. The residue obtained by concentrating the filtrate under reduced pressure and 1.78 g of a salt of benzyl trans-4-aminocyclohexanecarboxylate and p-toluenesulfonic acid were added to 20 ml of chloroform, and 950.4 mg of triethylamine was further added dropwise at 0°C. Add 200ml of ethyl acetate to this reaction mixture and add water.
Washed with 20 ml of 6% hydrochloric acid three times, 6 ml of 5% sodium bicarbonate solution three times, and saturated saline. After this washing, the organic layer was dried with Glauber's salt and concentrated under reduced pressure. The resinous residue was purified by silica gel chromatography (developing solvent: 30% acetone-n-hexane) to obtain 1.483 g of a colorless solid substance (f). The analysis results of the ^-1 HNMR spectrum of the obtained diamide (he) are shown below. ^-1H -NMR ( _CDCl3 ): δ1.17 (6H, d, J=6
Hz), 1-2.33 (9H, brm), 2.17 (6H, s,) 3.73
(1H, brs), 4.07 (1H, m), 5.13 (2H, s), 5.63
(2H, s), 6.43−6.77 (2H, brm), 7.35 (5H,
s). Add 6 ml of anhydrous THF to diamide (f) and add 1N at 0℃.
Deprotection was carried out by adding 2 ml of sodium hydroxide dropwise.
The reaction mixture was extracted by adding 150 ml of chloroform and 10 ml of water, and the organic layer was dried over Glauber's salt and concentrated under reduced pressure. The obtained crystalline residue was recrystallized from chloroform-methanol to obtain 503.6 mg of hydroxyamide (t). The analysis results of the ¹ HNMR spectrum of the obtained hydroxyamide (t) are shown below. ^1H -NMR ( _CDCl3 ): δ1.00-2.33 (9H, m),
1.15 (3H, d, J = 6Hz), 1.18 (3H, d, J =
6.7Hz), 3.33-4.33 (4H, m), 5.1 (2H, s),
5.17−5.50 (2H, m), 6.77−7.10 (2H, brm),
7.33 (5H, s). 20 ml of 80% ethanol-water and 200 mg of 10% Pt-C were added to 486.2 mg of hydroxyamide (t), and hydrogenated at normal pressure. The catalyst was filtered off, and the filtrate was concentrated under reduced pressure to obtain 382 mg of a colorless solid substance. 25 mg of the obtained carboxylic acid derivative was dissolved in 15 ml of isopropyl alcohol and 100 ml of n-hexane, and this was added to a column packed with aminopropyl silica with a diameter of 5 μm (ψ1
mm x 50 cm), and the stationary phase was ionically bonded to the substrate (silica) as expressed by formula (H) of the above reaction formula. Example 5 Application to optical resolution Using the chromatography stationary phase prepared in Example 4, β-hydroxycarboxylic acid amide having the following structural formula was separated in the same manner as in Example 3. The analysis results are shown in Table 2.

[Table] As is clear from Table 2, the stationary phase of the present invention separates various β-hydroxycarboxylic acid amides well. Example 6 Preparation of covalent stationary phase 1.24 g of monoamide (b) obtained in Example 1 and 528.2 mg of N-hydroxysuccinimide were added to 20 ml of anhydrous THF, and 1.0 g of DCC was dissolved in 1 ml of anhydrous THF at 0°C.
Add another g. The precipitated urea derivative is filtered off and washed with anhydrous THF. The filtrate is concentrated under reduced pressure, and 20 ml of chloroform is added to the resulting residue, and 1.0 g of aminopropyltriethoxysilane and 456 mg of triethylamine are added dropwise thereto. The reaction mixture is concentrated under reduced pressure, and the resulting residue is purified by silica gel chromatography (developing solvent: benzene). Add 1 g of silica gel and 20 ml of anhydrous toluene to 1.2 g of purified product (Li), and reflux the mixture for 24 hours. The slurry of silica gel is collected by filtration, washed with methanol, suspended in 5 ml of saturated ammonia-methanol, and left at 0°C for 8 hours. The obtained solid substance (N) is collected by filtration, washed with methanol, chloroform, and n-hexane, and dried. The stationary phase obtained is 900 mg. Example 7 Preparation of starting materials 10 ml of anhydrous methanol was added to 4.072 g of tartaric anhydride diacetate (a) under ice water cooling, and the mixture was allowed to stand for 1 hour. The reaction mixture was concentrated under reduced pressure, and the resulting crystalline residue was recrystallized from ethyl acetate-n-hexane. The yield of the obtained monoester (ru) was 3.71g.
It was hot. The analysis results of ¹ HNMR spectrum of this monoester were as follows. ¹ HNMR (CDCl ₃ ), δ: 2.176 (3H, s), 2.199
(3H, s), 3.792 (3H), 5.714−5.759 (2H, m). Example 8 Preparation of covalent stationary phase 1.516 g of the monoester obtained in Example 7 and 1.476 g of aminopropyl silica were added to 10 ml of anhydrous chloroform, and 2.5 ml of anhydrous methylene chloride was added at 0°C.
2.2 g of DCC dissolved in was added. Bring it up to room temperature,
100 ml of chloroform was added to the reaction mixture and centrifuged. The supernatant liquid was removed, and the silica derivative was collected by filtration, and then washed with chloroform, methanol, acetone, and n-hexane, and after drying, 1.37 g of a pale yellow solid substance (O) was obtained. The analysis results are shown below. IR (KBr): 1750cm ^-1 , 1660cm ^-1 , 1550cm ^-1 . 8 ml of 20% methylamine-methanol was added to 1.04 g of silica derivative (E), and the mixture was left at 0°C for 8 hours. The obtained solid substance was collected by filtration and washed with methanol, chloroform, and n-hexane. The yield of the chromatography stationary phase (wa) obtained after drying was 960 mg. The analysis results are shown below. IR (KBr): 1650cm ^-1 , 1550cm ^-1 .

Claims (1)

[Scope of Claims] 1. A stationary phase for chromatography chemically bonded to a substrate, characterized by having a chiral carbamoylhydroxycarboxylic acid amide residue represented by the following structural formula (I). (In the formula, R ₁ , R ₂ and R ₃ mean a hydrogen atom, an alkyl group or an aryl group, and n is an integer of 1 to 6.) 2 In formula (I), R ₁ , R ₂ and R ₃ are The stationary phase according to claim 1, which is a hydrogen atom, a linear or branched alkyl group having 1 to 6 carbon atoms, a phenyl group, a naphthyl group, or an anthryl group. 3 A chiral carbamoylhydroxycarboxylic acid derivative of the following structural formula (a) is condensed with an amino group chemically bonded to a substrate represented by the following structural formula (), and if necessary, the hydroxyl group of the derivative is protected before condensation. A method for producing a stationary phase for chromatography having a chiral carbamoylhydroxycarboxylic acid amide residue of the following structural formula (I) and chemically bonded to a substrate, which comprises deprotecting the compound after condensation. (In the formula, R ₁ , R ₂ and R ₃ mean a hydrogen atom, an alkyl group or an aryl group, and n is an integer from 1 to 6) (wherein R ₁ and R ₂ have the same meanings as above,
n is the same integer as above, and X means a group that activates a hydroxyl group, halogen, or carbonyl group) (In the formula, R ₃ means a hydrogen atom, an alkyl group or an aryl group, Car means a substrate, and R ₄ means a carbon chain or direct bond connecting the above amino group and the substrate) 4 Formula 4. The method according to claim 3, wherein when X in (a) is a hydroxyl group, a dehydration condensation agent is added during the condensation. 5 Neutralize the carboxyl group of the chiral carbamoylhydroxycarboxylic acid derivative of the following structural formula (a) with the amino group bonded to the substrate of the following structural formula (), and if necessary, remove the hydroxyl group of the derivative before neutralization. A method for producing a stationary phase for chromatography having a chiral carbamoylhydroxycarboxylic acid amide residue of the following structural formula (I) and chemically bonded to a substrate, which comprises protecting and deprotecting after neutralization. (In the formula, R ₁ , R ₂ and R ₃ mean a hydrogen atom, an alkyl group or an aryl group, and n is an integer from 1 to 6) (In the formula, R ₁ , R ₂ and R ₃ have the same meanings as above, n is the same integer as above, and R ₄ is a carbon chain or direct bond connecting adjacent carbon atoms and nitrogen atoms. ) (In the formula, R ₅ and R ₇ mean a hydrogen atom, an alkyl group, or an aryl group, and Car means a substrate) 6. A chiral carbamoylhydroxycarboxylic acid of the following structural formula (I), which is condensed with a hydroxyl group bonded to a substrate of formula (), and optionally protects the hydroxyl group of the derivative before condensation and deprotects it after condensation. A method for producing a stationary phase for chromatography having an amide residue and chemically bonded to a substrate. (In the formula, R ₁ , R ₂ and R ₃ mean a hydrogen atom, an alkyl group or an aryl group, and n is an integer from 1 to 6) (In the formula, R ₁ , R ₂ and R ₃ have the same meanings as above, n is the same integer as above, and R ₄ represents a carbon chain or direct bond connecting the adjacent nitrogen atom and silicon atom. and R ₈ , R ₉ , R ₁₀ are alkyl groups) HO-Car () (In the formula, Car means a substrate) A hydroxycarboxylic acid derivative is condensed with an amino group chemically bonded to a substrate represented by the following structural formula (), and then the alkoxycarbonyl group is replaced with a carbamoyl group,
A compound having a chiral carbamoylhydroxycarboxylic acid amide residue of the following structural formula (I) and chemically bonded to a substrate, which comprises protecting the hydroxyl group of the derivative before condensation and deprotecting it after condensation as necessary. Method for manufacturing stationary phases for chromatography. (In the formula, R ₁ , R ₂ and R ₃ mean a hydrogen atom, an alkyl group or an aryl group, and n is an integer from 1 to 6) (In the formula, n is the same integer as above, X means a group that activates a hydroxyl group, halogen or carbonyl group, and R ₁₁ means an alkyl group) (In the formula, R ₃ means a hydrogen atom, an alkyl group or an aryl group, Car means a substrate, and R ₄ means a carbon chain or direct bond connecting the above amino group and the substrate) 8 Formula 8. The method according to claim 7, wherein when X in (a) is a hydroxyl group, a dehydration condensation agent is added during the condensation. 9 A chiral hydroxycarboxylic acid derivative of the following structural formula (b) having an alkoxycarbonyl group is neutralized with an amino group bonded to a substrate of the following structural formula (V), and then the alkoxycarbonyl group is replaced with a carbamoyl group. , having a chiral carbamoylhydroxycarboxylic acid amide residue of the following structural formula (I), which comprises protecting the hydroxyl group of the derivative before neutralization and deprotecting it after neutralization, if necessary, and having a chemical compound on the substrate. Method for manufacturing a bonded stationary phase for chromatography. (In the formula, R ₁ , R ₂ and R ₃ mean a hydrogen atom, an alkyl group or an aryl group, and n is an integer from 1 to 6) (In the formula, n is the same integer as above, R ₃ has the same meaning as above, R ₄ means a carbon chain or direct bond connecting adjacent carbon atoms and nitrogen atoms, R ₁₁ is (means an alkyl group) (In the formula, R ₅ and R ₇ are hydrogen atoms, alkyl groups,
or an aryl group, and Car means a substrate) 10 The alkoxy group of the chiral trialkoxysilane derivative of the following structural formula (b) having an alkoxycarbonyl group is connected to the hydroxyl group bonded to the substrate of the following structural formula (). chiral carbamoyl hydroxy of the following structural formula (I), which consists of condensation, then substituting the alkoxycarbonyl group with a carbamoyl group, and optionally protecting the hydroxyl group of the derivative before condensation and deprotecting it after condensation. A method for producing a stationary phase for chromatography having a carboxylic acid amide residue and chemically bonded to a substrate. (In the formula, R ₁ , R ₂ and R ₃ mean a hydrogen atom, an alkyl group or an aryl group, and n is an integer from 1 to 6) (In the formula, R ₃ has the same meaning as above, R ₄ means a carbon chain or direct bond connecting the adjacent nitrogen atom and silicon atom, R ₈ , R ₉ , R ₁₀ and
R ₁₁ means an alkyl group) HO−Car () (in the formula, Car means a substrate)