Strong anion exchange composition and methods

• US 4,430,496 A
• Filed: 03/16/1982
• Issued: 02/07/1984
• Est. Priority Date: 01/17/1980
• Status: Expired due to Term

• Current Assignee: Varian Incorporated (Agilent Technologies, Inc.)
• Original Assignee: Varian Associates, Inc.
• Inventors: Abbott, Seth R.
• Primary Examiner(s): Moyer, Donald B.
• Assistant Examiner(s): Foley, Shawn P.
• Application Number:
  US06/359,173
• Time in Patent Office: 693 Days
• Field of Search: 536/27, 536/28, 536/29, 210/198.2, 210/635, 210/656, 556/425
• US Class Current: 536/26.73
• CPC Class Codes: B01J 20/264 derived from different types of monomers, e.g. linear or branched copolymers, block copolymers, graft copolymers
• CPC Class Codes: B01J 20/267 Cross-linked polymers
• CPC Class Codes: B01J 20/28004 Sorbent size or size distribution, e.g. particle size
• CPC Class Codes: B01J 20/28057 Surface area, e.g. B.E.T specific surface area
• CPC Class Codes: B01J 20/28083 being in the range 2-50 nm, i.e. mesopores
• CPC Class Codes: B01J 20/286 Phases chemically bonded to a substrate, e.g. to silica or to polymers
• CPC Class Codes: B01J 20/3204 Inorganic carriers, supports or substrates
• CPC Class Codes: B01J 20/321 consisting of a polymer obtained by reactions involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3212 consisting of a polymer obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3217 Resulting in a chemical bond between the coating or impregnating layer and the carrier, support or substrate, e.g. a covalent bond
• CPC Class Codes: B01J 20/3257 the functional group or the linking, spacer or anchoring group as a whole comprising at least one of the heteroatoms nitrogen, oxygen or sulfur together with at least one silicon atom, these atoms not being part of the carrier as such
• CPC Class Codes: B01J 2220/54 Sorbents specially adapted for analytical or investigative chromatography
• CPC Class Codes: B01J 2220/58 Use in a single column
• CPC Class Codes: B01J 41/05 in the strongly basic form
• CPC Class Codes: B01J 41/09 Organic material
• CPC Class Codes: B01J 41/10 Inorganic material
• CPC Class Codes: B01J 41/20 Anion exchangers for chromatographic processes

Abstract

A strong anion exchange composition useful in monophosphate nucleotide isomer separations is comprised of an inert porous particle having a silyl alkyl ammonium compound fixedly attached by a covalent silicon-oxygen-silicon bond to the surface thereof. A process for synthesizing the strong anion exchange composition in an aqueous solvent is disclosed together with methods for separation of monophosphate nucleotide isomers, highly phophorylated nucleotides and aldopentose carbohydrates using a column packed with this strong anion exchange composition.

Claims

1. A method for the separation of components contained in a biological composition, the steps of the method comprising:

(a) packing a column with a strong anion exchange composition comprising an inert porous particle having a particle size of about 3 to about 20 microns which has covalently attached to the silica surface a silyl alkyl ammonium compound having the general formula;

##STR4## wherein R is methyl or ethyl;

n is an integer ranging from 2 to 5; and

X is an exchangeable anion;

(b) passing said biological composition through said column; and

,

(c) recovering the separated components.

2. The method of claim 1 wherein said column is a high-pressure liquid chromatography column.

3. The method of claim 2 wherein the particle of the strong anion exchange composition is selected from a group consisting of microparticulate silica, alumina, a cross-linked dextran, and a cross-linked polystyrene-divinylbenzene resin.

4. The method of claim 3 wherein the particle of which said strong anion exchange composition is comprised is microparticulate silica.

5. The method of claim 4 wherein said silica is a high-pressure liquid chromatography grade silica gel.

6. The method of claim 2 wherein the said biological composition separated by said strong anion exchange composition is comprised of members of a group consisting of nucleic acid components, amino acids, and dicarboxylic acids.

7. The method of claim 6 wherein the said biological composition separated by said strong anion exchange composition is a mixture of nucleic acid components.

8. The method of claim 1 wherein said particle size is about 3-10 microns.

9. The method of claim 1 wherein n is 3.

10. The method of claim 9 wherein R is methyl.

11. The method of claim 1 wherein said ammonium compound is N-trimethoxy silyl-propyl-N,N,N-trimethylammonium chloride.

12. A liquid chromatographic column comprising an elongated tube, a strong anion exchange composition packed into said tube, said strong anion exchange composition comprising an inert porous particle having a size of about 5 to 20 microns and having attached to the surface of said inert porous particle by covalent bonding a silyl alkyl ammonium compound having the general formula:

##STR5## wherein R is methyl or ethyl;

n is an integer ranging from 2 to 5; and

X is an exchangeable anion, said strong anion exchange composition being capable of simultaneous separation of mixtures of polyfunctional compounds and isomers thereof.

13. The column of claim 12 wherein said column is a high-pressure liquid chromatography column.

14. The column of claim 13 wherein the particle of the strong anion exchange composition is selected from a group consisting of microparticulate silica, alumina, a cross-linked dextran, and a cross-linked polystyrene-divinylbenzene resin.

15. The column of claim 14 wherein the particle of which said strong anion exchange composition is comprised is microparticulate silica.

16. The column of claim 15 wherein said silica is a high-pressure liquid chromatography grade silica gel.

17. The column of claim 14 wherein said particle size is about 5-10 microns.

18. The column of claim 12 wherein n is 3 and R is methyl.

19. The column of claim 12 wherein said ammonium compound is N-trimethoxy silyl-propyl-N,N,N-trimethylammonium chloride.

20. A method for performing liquid chromatography comprising placing a solution of a composition comprising a mixture of polyfunctional compounds and isomers thereof on the head of a chromatographic column, flowing a solvent system through said chromatographic column, wherein said chromatographic column is packed with a packing material comprising a strong anion exchange composition, said strong anion exchange composition comprising an inert porous particle having a size from about 3 to 20 microns which has covalently attached to its surface a silyl alkyl ammonium compound having the general formula:

##STR6## wherein R is methyl or ethyl;

n is an integer ranging from 2 to 5; and

X is an exchangeable anion;

wherein said strong anion exchange composition is capable of simultaneous separation of mixtures of polyfunctional compounds and isomers thereof.

21. The method of claim 20 wherein said column is a high-pressure liquid chromatography column.

22. The method of claim 21 wherein the particle of the strong anion exchange composition is selected from a group consisting of microparticulate silica, alumina, a cross-linked dextran, and a cross-linked polystyrene-divinylbenzene resin.

23. The method of claim 22 wherein the particle of said strong anion exchange composition is comprised is microparticulate silica.

24. The method of claim 23 wherein said silica is a high-pressure liquid chromatography grade silica gel.

25. A method for the separation of nucleotide isomers on a chromatographic column, the steps of the method comprising:

(a) packing a column with a strong anion exchange composition comprising a microparticulate silica having a particle size of about 3 to about 20 microns which has covalently attached to the silica surface a silyl alkyl ammonium compound having the general formula;

##STR7## wherein R is methyl or ethyl;

n is an integer ranging from 2 to 5; and

X is an exchangeable anion;

(b) passing said nucleotide isomer composition through said chromatographic column; and

(c) recovering the separated components.

26. The method of claim 25 wherein the said nucleotide isomers separated by said strong anion exchange composition are comprised of members of a group consisting of nucleic acid components, amino acids, and dicarboxylic acids.

27. The method of claim 26 wherein the said nucleotide isomers comprise a mixture of nucleic acid components.

28. The method of claim 25 wherein n is 3 and R is methyl.

29. The method of claim 25 wherein said ammonium compound is N-trimethoxy silyl-propyl-N,N,N-trimethylammonium chloride.