JPS60193929A - Separation agent - Google Patents

Abstract

PURPOSE:To provide a separating agent suitable for the separation of optical isomers, by preparing a poly-alpha-amino acid having a polymerization degree of within a specific range and derived from monomers such as alanine, valine, etc., and supporting the amino acid to a carrier such as a porous organic carrier. CONSTITUTION:A poly-alpha-amino acid having a polymerization degree of 2- 1,000,000, preferably 4-100,000 (e.g. a polymer derived from the monomers such as leucine, isoleucine, threonine, serine, methionine, phenylalanine, glutamic acid, their ether, ester, amide, urethane, etc.) is supported on a carrier, preferably a porous organic carrier such as polystyrene, polyacrylamide, polyacrylate, etc. The separation agent is preferably used especially in a liquid chromatography or thin-layer chromatography.

Description

DETAILED DESCRIPTION OF THE INVENTION (1) Industrial Application Field The present invention relates to the use of poly-α-amino acids as separation agents for substances.

The substances to be separated are not only ordinary low-molecular compounds, but also optical isomers, which have traditionally been very difficult to separate directly.

Separation of optical isomers is often necessary, for example, in the fields of medicine, agrochemicals, etc., in order to prevent drug damage and improve drug efficacy per unit amount used. Expanded priority crystallization and diastereomer methods have conventionally been used to separate optical isomers, but these methods limit the types of optical isomers that can be separated, and require a long time. Often inefficient.

On the other hand, since separation by chromatography, 41FKi body chromatography, and thin layer chromatography is simple, there has been a strong desire to develop efficient separation agents for use in these methods.

(2) Prior art An example of separating optical isomers by gas-liquid partition chromatography using various monomeric α-amino acid derivatives and low molecular weight peptide derivatives as a stationary phase is Tetra hedro.
n Letters, 10.1009 (1966),
Special Publication No. 44-25957, Special Publication No. 52-136188,
Ochromatographia, 9.

351 (1976), Ochromatography
a, 10, 444 (1977), but since they all use low molecular weight substances, their solubility is high and it is difficult to use them in the liquid chromatography method or thin layer chromatography method. In addition, wool keratin and b9vin ser
There are known examples of separation of optical isomers using a protein such as umalbumin, but all four have random polymer structures and the details are not yet clear.

(3) Constituent composition of the invention In the general formula R', R represents a terminal amino group and a carboxyl group. Natural or synthetic amino acids forming the formula H2N-0-000H, and ethers, esters, amides, urethanes, imines, and imides derived therefrom. , acid anhydrides, carboxylates, mineral acid salts.

Amine salts are residues formed from metal salts.

R' is hydrogen, a hydroxyl group, or a hydroxyl group, a highly derived ether, an ester, or a urethane.

These may be homopolymers with the same R or R', copolymers or graft polymers with different R or R', or a mixture of two or more types of poly-α-amino acids, or poly-α- Amino acids and other resins may be mixed.

When the poly-α-amino acid is a copolymer, it is necessary that one of the amino acids has a molar ratio of at least 50% or that the copolymer has at least three components.

m and n represent the degree of polymerization, each from 2 to 1.000
,000, preferably from 4 to 100,000.
The optically active constituent monomer units of α-amino acids are:
Alanine, valine, leucine, inleucine, threonine, serine, methionine, phenylalanine, florine.

Trizutophan, histidine, glutamic acid.

Examples include glutamine, aspartic acid, asparagine, arginine, lysine, ornithine, and ethers, esters, amides, urethanes, imines, imides, acid anhydrides, and salts derived from these.

Further, in order to improve the pressure resistance of the separation agent, prevent swelling and shrinkage due to solvent substitution, and increase the number of theoretical plates, it is preferable that the poly-α-amino acid is retained on a carrier.

The carrier may be a porous organic carrier or a porous inorganic carrier, preferably a porous inorganic carrier. Suitable porous organic carriers include polymeric materials such as polystyrene, polyacrylamide, polyacrylate, and the like. Suitable porous inorganic carriers include synthetic or natural materials such as silica, alumina, magnesia, titanium oxide, glass, silicates, and kaolin, which can be used to improve their affinity with poly-α-amino acids. Surface treatment may also be performed. Examples of surface treatment methods include silanization using an organic silane compound and surface treatment using plasma polymerization.

The suitable size of the carrier varies depending on the size of the column or plate used, but is generally 1 pm to 10 fins, preferably 1 pm to 300 pm. Preferably, the carrier is porous, with an average pore size of 500=OOA. The amount of poly-α-amino acid retained is 1 to 1 to the carrier.
00% by weight, preferably 5 to 50% by weight.

A method for retaining poly-α-amino acids on a carrier may be a chemical method or a physical method. Physical methods include a method in which the poly-α-amino acid is dissolved in a soluble solvent, mixed well with a carrier, and the solvent is distilled off by air flow under reduced pressure or heating.

After dissolving the poly-α-amino acid in a soluble solvent and thoroughly mixing it with the carrier, stirring it into a liquid that is incompatible with the solvent,
There is also a method of dispersing and diffusing the solvent.

By adding a small amount of solvent to the poly-α-amino acid retained on the carrier in this way, the poly-α-amino acid is temporarily swollen or dissolved, and by distilling off the solvent again, the retention state and thus the separation ability can be changed. is possible.

(4) Effects of the Invention Chromatography is suitable for using the separating agent of the present invention containing poly-α-amino acids as a main component for the purpose of separating compounds. As a chromatographic method, a liquid chromatographic method or a thin layer four-layer method is preferable.

When used as a liquid chromatography method, a column is filled with poly-α-amino acids supported on a carrier.

In addition, when performing thin layer chromatography, 0.1 μm to 0.1 μm
A layer having a thickness of 0.1 nm to 100 nm, consisting of the separation agent of the present invention consisting of particles of about n size and, if necessary, a small amount of a binder, may be formed on the support plate.

The separating agent containing poly-α-amino acids as a main component of the present invention is effective for separating compounds, and is particularly effective for resolving nine optical isomers, which are conventionally very difficult to separate. The optical isomers to be separated are compounds with an asymmetric center or molecularly asymmetric compounds, and one of the optical isomers is strongly retained by poly-α-amino acids.

The present invention will be described in detail below with reference to Examples.

The present invention is not limited to these examples. The definitions of terms used in the examples are as follows.

For high performance liquid chromatography, JASCO Corporation's TR
Engineered ROTORIF was used. The detectors include an ultraviolet absorption measuring device UV FiO100■ manufactured by JASCO Corporation and a polarimeter D-P 1810 (7 RU: 5 BEN 04G manufactured by JASCO Corporation).
(id,) 俤) was used for detection at a wavelength of 356 nm.

Synthesis method Poly T-benzyru-glutamate (Sigma, molecular weight 50,000) o, 6 oat chloro, form 1011
Silica gel dissolved in Ll and treated with 3-7 minoprobyltriethoxysilane (Merck, Li0hro
spher 8 engineering 4000) 3. ost and evaporate the solvent.

(The solution was divided into two batches of 51 kg each.) Example This packing material was packed into a column with a length of 25 os and an inner diameter of 0.46 m by the slurry method, and hexane-2-panol (q a : 2 ), was used as the eluent at a flow rate of 0.5
The resolution of racemates A and B was carried out at su/min and 25°C. As a result, A leaked out after a retention time of 15.8 minutes, and B'' flowed out after a retention time of 36.5 minutes, and when the front end was separated and the optical rotation was measured at a wavelength of 15 nm, both showed positive (+) optical rotation. Indicated.

A B Device TR↓ROTAR-II, UVIDIC-100-I
I Applicant's agent Kaoru Furuya Procedural amendment (method) ■, Indication of the case Japanese Patent Application No. 51-47893 2, Title of the invention Separation agent 3, Person making the amendment Relationship with the case Patent applicant (29, 0) Daicel Chemical Industries, Ltd. 4, Agent: Nakai Building, 1-3 Nihonbashi Yokoyama-cho, Chuo-ku, Tokyo June 1982
May 26th (shipment date) 6. Specifications subject to amendment 7. Contents of amendment

Claims (1)

[Claims] Poly-α- whose main component is a polymerization degree of 2 to 1,000,000
A separation agent made by supporting amino acids on a carrier.