Sample preparation for high throughput purification

• US 6,387,273 B1
• Filed: 08/25/2000
• Issued: 05/14/2002
• Est. Priority Date: 08/27/1999
• Status: Expired due to Fees

• Current Assignee: Scynexis Incorporated
• Original Assignee: SCYNEXIS Chemistry & Automation, Inc.
• Inventors: Abedi, Jaleh
• Primary Examiner(s): Therkorn, Ernest G.
• Application Number:
  US09/645,348
• Time in Patent Office: 627 Days
• Field of Search: 210/635, 210/656, 210/659, 210/143, 210/198.2, 436/161
• US Class Current: 506/30
• CPC Class Codes: B01D 15/12 relating to the preparation of the feed
• CPC Class Codes: B01D 15/16 relating to the conditioning of the fluid carrier
• CPC Class Codes: B01D 15/163 Pressure or speed conditioning
• CPC Class Codes: B01D 15/203 Equilibration or regeneration
• CPC Class Codes: B01D 15/424 Elution mode
• CPC Class Codes: B01J 2219/00452 Means for the recovery of reactants or products
• CPC Class Codes: B01J 2219/00587 High throughput processes
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01J 2219/0072 Organic compounds
• CPC Class Codes: B01J 2219/00759 Purification of compounds synthesised

Abstract

An HPLC method for purifying and/or characterizing large numbers of related compounds, for example, combinatorial and lead generation libraries, is disclosed. The compounds are purified on a semi-preparative or preparative scale without having to first remove the solvent from the vessel in which the compounds were prepared. This can be accomplished by equilibrating the HPLC column with a first mobile phase such that, when the sample is added to the column in the solvent in which it is prepared, the sample does not significantly elute off of the column in the void volume, and the solvent in which the compound is prepared does elute off of the column in the void volume or shortly thereafter. In addition, the solvent power can be adjusted such that the mobile phase is not a strong solvent for the compound of interest. The size of the void volume can also be decreased, for example, using a relatively short column and asorbent with a particle size of about 5 microns or less, with suitable inlet and outlet tubing to handle the increased pressure. The column can be saturated with a weak solvent prior to sample injection, for example, by increasing the flow rate of a mobile phase through the HPLC column.

Claims

1. A method for preparing samples for HPLC purification of a library of compounds without having to remove a first solvent prior to placing compounds on a column, comprising:

a) selecting a first mobile phase such that when the void volume of the mobile phase is combined with the amount of solvent in the vessel used to prepare the compound of interest, the resulting solvent system does not cause the compound of interest to significantly elute off of the column,

b) conditioning the column with a selected mobile phase,

c) applying the compound dissolved in the solvent used to prepare the compound to the column, and

d) eluting the solvent used to prepare the compound of interest from the column.

2. The method of claim 1, further comprising applying a second mobile phase to the column to elute the organic compound from the column.

3. The method of claim 1, further comprising regeneration-washing the column after the compound of interest has eluted.

4. The method of claim 1, further comprising increasing the flow rate of the mobile phase relative to that used in a conventional HPLC such that the compound does not significantly elute off of the column in the void volume or cause a blockage in the HPLC system.

5. The method of claim 4, further comprising increasing the tubing size of the HPLC relative to that used in a conventional preparative HPLC to compensate for the increased column back pressure due to the increased flow rate.

6. The method of claim 1, further comprising decreasing the particle size of the sorbent relative to that used in a conventional preparative HPLC to slow down the elution of the compound of interest from the HPLC column.

7. The method of claim 6, further comprising increasing the tubing size of the HPLC relative to that used in a conventional preparative HPLC to compensate for the increased column back pressure due to the relatively small sorbent particle size.

8. The method of claim 1, further comprising performing route scouting on a representative sample of a library of compounds to determine appropriate conditions for eluting the solvent used to prepare the compounds from the HPLC column without significantly eluting the compounds of interest from the HPLC column.