Additives for reversed-phase HPLC mobile phases
AssigneeAgilent Technologies, Inc.
InventorsBrian Bidlingmeyer, Qunjie Wang
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Abstract
The present invention provides silica-based reversed-phase HPLC methods that lead to higher retention of the analytes in the column and longer column lifetimes than usually observed under medium to high pH aqueous mobile phase conditions. The inventive methods comprise eluting the HPLC column using an aqueous mobile phase comprising at least one fluorinated additive. Preferred additives are polyfluorinated alcohols such as 2,2,2-trifluoroethanol and 1,1,1,3,3,3-hexafluoroisopropanol. The methods of the present invention may be used for analyzing, separating, purifying, and/or isolating small organic molecules, natural products, as well as biomolecules such as polypeptides, oligonucleotides and polynucleotides (e.g., DNA fragments).
Claims26
A method for increasing column lifetime for a HPLC column packed with a superficially porous silica-based reversed-phase support, the method comprising:
- eluting the HPLC column, which is packed with a superficially porous silica-based reversed-phase support and loaded with a sample mixture comprising at least two components, with an aqueous mobile phase comprising between 0.5 and 10% by volume of at least one neutral, polar, fluorinated organic compound;
- wherein the presence of the neutral, polar, fluorinated organic compound in the aqueous mobile phase leads to an increased column lifetime, as compared with the lifetime observed in the absence of the neutral, polar, fluorinated organic compound, all other conditions being equal.
A method for improving resolution of sample mixture components by increasing retention of at least one component of a sample mixture comprising at least two components, the method comprising:
- eluting a HPLC column, which is packed with a superficially porous silica-based reversed-phase support and loaded with a sample mixture comprising at least two components, with an aqueous mobile phase comprising:
- between 0.5 to 10% by volume of an additive comprising at least one neutral, polar, fluorinated organic compound;
- and
- wherein the presence of the neutral, polar, fluorinated organic compound in the aqueous mobile phase leads to a higher retention of at least one component of the mixture as compared with the retention observed for the same component of the mixture in the absence of the neutral, polar, fluorinated organic compound, all other conditions being equal.
Dependencies are inferred from the claim text.