摘要 |
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. |