Purification of substances by reaction affinity chromatography

• US 7,094,351 B2
• Filed: 10/12/2001
• Issued: 08/22/2006
• Est. Priority Date: 10/12/2000
• Status: Expired due to Fees

• Current Assignee: Robert C. Corcoran
• Original Assignee: Robert C. Corcoran
• Inventors: Corcoran, Robert C.
• Primary Examiner(s): Therkorn, Ernest G.
• Application Number:
  US09/975,258
• Publication Number:
  US 20020100727A1
• Time in Patent Office: 1,775 Days
• Field of Search: 210/635, 210/656, 210/659, 210/198.2, 436/161
• US Class Current: 210/635
• CPC Class Codes: B01D 15/3804 Affinity chromatography
• CPC Class Codes: B01D 15/3857 Reaction chromatography
• CPC Class Codes: B01J 20/281 Sorbents specially adapted for preparative, analytical or investigative chromatography
• CPC Class Codes: B01J 20/286 Phases chemically bonded to a substrate, e.g. to silica or to polymers
• CPC Class Codes: B01J 20/32 Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
• 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 20/3219 involving a particular spacer or linking group, e.g. for attaching an active group
• CPC Class Codes: B01J 20/3242 Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
• CPC Class Codes: B01J 20/3244 Non-macromolecular compounds
• CPC Class Codes: B01J 20/3248 the functional group or the linking, spacer or anchoring group as a whole comprising at least one type of heteroatom selected from a nitrogen, oxygen or sulfur, these atoms not being part of the carrier as such
• CPC Class Codes: B01J 20/3251 comprising at least two different types of heteroatoms selected from nitrogen, oxygen or sulphur
• CPC Class Codes: B01J 20/3253 comprising a cyclic structure not containing any of the heteroatoms nitrogen, oxygen or sulfur, e.g. aromatic structures
• CPC Class Codes: B01J 20/3255 comprising a cyclic structure containing at least one of the heteroatoms nitrogen, oxygen or sulfur, e.g. heterocyclic or heteroaromatic structures
• CPC Class Codes: B01J 2220/54 Sorbents specially adapted for analytical or investigative chromatography

Abstract

The invention relates to the separation of at least one target from a sample composition using a naturally reversible reaction comprising the formation of at least one covalent bond. One embodiment of the invention is a chromatographic method in which the affinity of a stationary phase for at least one target is based on the equilibrium constant of such a naturally reversible reaction between the stationary phase and the target.

Claims

1. A method of separating a target from a sample composition containing the target, comprising the following steps:

(a) contacting the sample composition with a reactive affinity molecule attached to a phase separating group, the reactive affinity molecule comprising a reactive functional group, and the reactive affinity molecule reacting with the target to farm an adduct by forming a covalent bond between the target and the reactive functional group, where the reaction forming the adduct is reversible under the conditions of the contacting without the addition of a reagent acting at the covalent bond, to reform the target and reactive affinity molecule

(b) separating the adduct from the sample composition;

(c) contacting the adduct with an eluent to regenerate the target and the reactive affinity molecule; and

(d) separating the target from the reactive affinity molecule.

2. The method of claim 1 where the reactive functional group comprises a group chosen from N═

N, C═

C, C═

O, N═

O, C═

N, C═

S, and C≡

C.

3. The method of claim 1 where the target comprises a 1,3-diene and the reactive functional group comprises a nitroso group.

4. The method of claim 1 where the target comprises a hydroxy, mercapto, or amino group and the reactive functional group comprises an aldehyde group.

5. The method of claim 1 where the target comprises a hydroxy, mercapto, or amino group and the reactive functional group comprises an alkene substituted by an anion stabilizing group.

6. The method of claim 1 where the target comprises a hydroxy, mercapto, or amino group and the reactive functional group comprises a ketone.

7. The method of claim 1 where the target comprises a hydroxy, mercapto, or amino group and the reactive functional group comprises an iminium group.

8. The method of claim 1 where the target comprises a hydroxy, mercapto, or amino group and the reactive functional group comprises a carboxylate ester.

9. The method of claim 1 where th reactive affinity molecule is attached to the phase separating group by a covalent bond, chemisorption, or ion-pairing.

10. The method of claim 1 where the phase separating group is a solid.

11. The method of claim 10 where the solid is chosen, from polymers, silica, alumina, and carbon.

12. The method of claim 11 where the solid is a polymer.

13. The method of claim 12 where the solid is a macroreticular polymer.

14. The method of claim 12 where the solid is chosen from polyethers, polyamides, polyesters, and polyenes.

15. The method of claim 12 where the solid is chosen from polyacrylates and polystyrene.

16. The method of claim 10 where the solid is a stationary phase of a chromatographic column.

17. The method of claim 1 where the phase separating group comprises a group that imparts to the adduct solubility in a liquid.

18. The method of claim 17 where the group that imparts solubility in a liquid is a water solubilizing group.

19. The method of claim 18 where the group that imparts solubility in a liquid is chosen from sulfate, sulfonate, phosphate, phosphonate, carboxylate, ammonium, phosphonium, sulfonium, and guanidinium ions, polyhydric alcohols, and polyethers.

20. The method of claim 17 where the group that imparts solubility in a liquid imparts solubility in a water immiscible phase.

21. The method of claim 1 where the reactivity molecule further comprises a reactivity modifier group.

22. The method of claim 21 where the reactivity modified group comprises an acidic group chosen from carboxylic adds, phenols, ammonium cations, phosphates, phosphate esters, phosphonates, and phosphonate esters.

23. The method of claim 21 where the reactivity modifier group comprises a basic group chosen from amines, heteroaryl amines, carboxylates, phenolates, phosphate anions, and phosphonate anions.

24. The method of claim 21 where the reactivity modifier group is chosen from halogens, nitro groups, cyano groups, hydroxyl groups, alkoxy groups, fluoroalkyl groups, perfluoroalkyl groups, nitrile groups, carboxyl groups, carboxylic ester groups, amide groups, sulfoxide groups, sulfone groups, carbonyl groups, and ammonium groups.

25. The method of claim 21 where the reactivity modifier group is chosen from hydroxyl groups, amine groups, monoalkylamine groups, dialkylamine groups, and alkoxy groups.

26. The method of claim 21 where the reactivity modifier group alters a property of the reactive functional group chosen from electronic characteristics, steric availability, and chirality.

27. The method of claim 21 where the eluent changes the equilibrium constant of the reaction by modifying the reactivity modifier group.

28. The method of claim 1 where the reactive affinity molecule further comprises a framework group.

29. The method of claim 28 where the framework group comprises a group chosen from alkyl groups, aryl groups, and heteroaryl groups.

30. The method of claim 1, where the reactive affinity molecule comprises a group of formula

31. The method of claim 30 where the reactive affinity molecule comprises a group chosen from R_1a(R_1b)C═

C(R_2a)R_2b, R_1a(R_1b)C═

O R_1a(R_1b)C═

N—

R_2a, and R_1a—

N═

O.

32. The method of claim 1 where the reactive affinity molecule comprises a group of formula

33. The method of claim 1 where the reactive affinity molecule comprises a group of formula

34. The method of claim 33 where Z comprises a group chosen from nitro, ketone, ester, amide, sulfoxide, sulfone, nitrile, iminium, and phosphonium groups.

35. The method of claim 1, where the reactive affinity molecule comprises a group of formula

36. The method of claim 1, where the reaction comprises the formation of at least two covalent bonds.

37. The method of claim 1 where the eluent is chosen from water, alcohols, hydrocarbons, and ethers.

38. The method of claim 37 where the eluent is chosen from methanol, ethanol, propanol, isopropanol and butanol.

39. The method of claim 37, where the eluent is chosen from benzene, toluene, xylenes, mesitylenes, hexanes, heptanes, octanes and nonanes.

40. The method of claim 37, where the eluent is chosen from propyl ethers and butyl ethers.

41. The method of claim 1, where the eluent has a property chosen from polarity, temperature, and pH that is different from the corresponding property of the sample composition.

42. The method of claim 41 where the eluent has a temperature that is different from the temperature of the sample composition.

43. The method of claim 1 where the half-life of the reaction is about 4 hours or less at 25°

C. under the contacting conditions.

44. The method of claim 1 where the step of contacting the adduct with the eluent also separates the target from the reactive affinity molecule.

45. The method of claim 1 where the target is chosen from ergosterol, thebaine, and vitamin D.

46. A method for isolating thebaine from a sample composition, comprising reversibly separating the thebaine from the sample composition by reverse affinity chromatography using with a reactive affinity molecule of the formula: